Compounds and Compositions for the Detection and Treatment of Alzheimer&#39;s Disease and Related Disorders

ABSTRACT

One aspect of the present invention relates to compounds, compositions and methods for diagnosis and/or treatment of a subject suffering from an amyloidosis-associated pathological condition. In certain embodiments, the imaging and/or therapeutic agents of the instant invention may be administered to a subject for identification and/or treatment of amyloid deposits. A specific imaging method detects amyloid deposits by administering the imaging agent to the subject and detecting the spatial distribution of the agent. Differential accumulation of the agent is indicative of AD or an amyloidosis-associated pathological condition and can be monitored by using a PET or SPECT camera.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.12/933,139, now U.S. Pat. No. 8,450,466, issued May 28, 2013, which isthe U.S. National Stage of International Patent Application No.PCT/US2009/037928, filed Mar. 23, 2009, which claims the benefit ofpriority to U.S. Provisional Patent Application Ser. No. 61/038,571filed Mar. 21, 2008, each of which is hereby incorporated by referencein its entirety.

BACKGROUND

Millions of Americans suffer from dementia and other cognitive deficitsas a result of Alzheimer's disease (AD), a neurodegenerative disease.Due to its occurrence in the brain, it is difficult to diagnose thecondition and to determine its cause without dangerous brain biopsy.Scientists believe that as many as 4.5 million Americans suffer from AD.AD usually begins after age 60 and its risk goes up with age. The causeof AD is unknown and, at present, no cure has been found.

AD can only be definitely confirmed after an autopsy, which preventsearly accurate diagnosis and treatment of the condition.Neuropathologically, AD is characterized by the presence of neuriticplaques, neurofibrillary tangles and neuronal loss. See Mann. Mech.Ageing Dev. 1985, 3(1), 213. Doctors can diagnose AD correctly up to 90percent using several tools to diagnose “probable” AD, namely, (1)questions about the person's general health, past medical problems, andability to carry out daily activities; (2) tests of memory, problemsolving, attention, counting, and language; (3) medical tests, such astests of blood, urine, or spinal fluid; and (4) brain scans.

Postmortem brain tissues of AD victims show the presence of amyloidcores of neuritic plaques that are composed of amyloid β-(Aβ-) proteinbeing predominantly arranged in beta-pleated sheet configuration. See J.Biol. Chem. 1992, 267(24), 17082; and Proc. Natl. Acad. Sci., USA 1986,83(2), 503.

Deposition of amyloid β-(A β-) protein occurs, however, not only inindividuals that have AD, but it also frequent among individuals who areundergoing the aging process. Thus, it is very critical to distinguishthe AD production due to the normal aging process or to AD or otherdementia-causing diseases such as DLB dementia associated with LouisBody. In the normal aging process, non-compact or diffuse amyloidplaques containing less fibrillar AP are deposited primarily in thebrain. In contrast, AD patients have brains that are characterized by anunanatomically widespread process of amyloid deposition and neuriteplaque formation containing dense amyloid fibrils.

Clinical tests to determine the onset of AD and its progression are notpresently sensitive and several agents are reported as potential PET andSPECT imaging tracers. Some of the developmental research on imagingagents useful for the diagnosis of AD and other related diseases arediscussed below.

U.S. Patent Publication Application No. 2006/0018825 A1, assigned to BFResearch Institute, hereby incorporated by reference, describes a seriesof BF compounds or a salt or solvate thereof that can be used as a probefor the imaging and diagnosis of diseases in which amyloid P-proteinaccumulates. These compounds have high specificity for diffuse plaquesand act as early indicators of AD. In addition, they have rapidclearance from the brain.

Okamura et al. (in J. Neurosci. 2004, 24(10), 2535) describes a labeledsterylbenzoxazole derivative compound, ¹⁸F-radiolabeled6-(2-fluoroethoxy)-2-[2-(4-methylaminophenyl)ethenyl]-benzoxazole(BF-168), that demonstrated abundant initial brain uptake (3.9% injecteddose/gm at 2 min after injection) and fast clearance (t_(1/2)=24.7 min)after intravenous (iv) administration in normal mice. In addition,autoradiograms of brain sections from APP23 transgenic mice at 180 minafter iv injection of ¹⁸F-radiolabeled BF-168 showed selective labelingof brain amyloid deposits with little non-specific binding.

More recently, Kudo et al. (in J. Nucl. Med. 2007, 48553) havedemonstrated the use of a novel compound, F-18 labeled2-(2-[2-diethylaminothiazol-5-yl]-ethenyl)-6-(2-[fluoro])ethoxybenzoxazol(eB F-227) as a promising PET probe for in vivo detection of denseamyloid deposits in AD patients.

U.S. Pat. Nos. 6,001,331 and 6,696,039 B2, issued Dec. 14, 1999 and Feb.24, 2004, respectively, hereby incorporated by reference, describe theuse of several radiolabeled benzothiazole compounds for imaging amyloiddeposits.

U.S. Pat. Nos. 6,168,776 and 6,133,259, issued Jan. 2, 2001 and Oct. 17,2000, respectively, hereby incorporated by describe amyloid-bindingcompounds such as Chrysamine G and their use in identifying AD in vivoand other pathological conditions characterized by amyloidosis.

One promising amyloid imaging agent is an analogue of thioflavin T, alsoknown as the Pittsburgh Compound-B or “PIB compound.” PIB is also knownas [N-methyl-(¹¹C)]-2-(4′-methylaminophenyl)-6-hydroxybenzothiazole (or[¹¹C]6-OH-BTA-I). PET imaging with ¹¹C-PIB can discriminate AD fromfrontotemporal lobar degeneration (FTLD). See J. Med. Chem. 2003,46(13), 2740; and Neurology 2007, 68, 1205. However, use of a C-11labeled tracer limits imaging to medical centers with a cyclotron.

It is well known that 2,8-diazaspiro[4,5]decane-1,3-dione (RS-86)derivatives are active and centrally effective muscarinic cholinergicagonists, with analgesic and sedative properties in animals when givenorally. In addition, it has been shown that the C-11 radiolabeledversion of 2,8-diazaspiro[4,5]decane-1,3-dione can be used as a tracer,though the reported study describes brain distribution results with avery low specific activity. Further, the biodistribution of the C-11radiolabeled 2,8-diazaspiro[4,5]decane-1,3-dione in rats, as a functionof time, showed that the initial brain uptake was about 1.1%, with highconcentrations of percent dose per gram in areas rich with muscarinicreceptors such as caudate, putamen and thalamus. However, as discussedabove with respect to ¹¹C-PIB, the utility of a C-11 labeled tracer islimited to medical centers with a cyclotron.

Accordingly, there is a need to provide compounds and methods forimaging and treating AD and amyloidosis-associated pathologicalconditions that are easily available and cost effective. There is acontinuing need to seek novel imaging tracers that are accurate and usedin early detection of AD and other related pathological conditions.

SUMMARY

One aspect of the present invention relates to compounds, compositionsand methods for diagnosis and/or treatment of a subject suffering froman amyloidosis-associated pathological condition. In certainembodiments, the imaging and/or therapeutic agents of the instantinvention may be administered to a subject for identification and/ortreatment of amyloid deposits. A specific imaging method detects amyloiddeposits by administering the imaging agent to the subject and detectingthe spatial distribution of the agent. Differential accumulation of theagent is indicative of AD or an amyloidosis-associated pathologicalcondition and can be monitored by using a PET or SPECT camera.

One aspect of the invention relates to the preparation of fluorinated orradiofluorinated 2,8-diazaspiro[4,5]decane-1,3-diones, and theirdiagnostic and/or therapeutic use in brain disorders associated withaging, such as, senile dementia and Alzheimer's disease.

Another aspect of the invention relates to the preparation offluorinated or radiofluorinated inositols (such as1-deoxy-1-fluoro-scyllo-inositol and 1-deoxy-1-fluoro-myo-inositol), andtheir diagnostic and/or therapeutic use in brain disorders associatedwith aging, such as, senile dementia and Alzheimer's disease.

Another aspect of the invention relates to compounds which are acombination of an inositol and a 2,8-diazaspiro[4,5]decane-1,3-dione(e.g., wherein the inositol is covalently linked by a cross-linker tothe 2,8-diazaspiro[4,5]decane-1,3-dione), and their diagnostic and/ortherapeutic use in brain disorders associated with aging, such as,senile dementia and Alzheimer's disease.

Another aspect of the invention relates to optionally fluorinated orradiofluorinated combinations of 2,8-diazaspiro[4,5]decane-1,3-dionesand D-glucose (e.g. [F-18]-2-fluoro-2-deoxy-D-glucose), and theirdiagnostic and/or therapeutic use in brain disorders associated withaging, such as, senile dementia and Alzheimer's disease. In certainembodiments, the 2,8-diazaspiro[4,5]decane-1,3-dione improves theselectivity and/or uptake of the glucose.

Another aspect of the invention relates to the optionally fluorinated orradiofluorinated combination of an inositol with D-glucose (e.g.[F-18]-2-fluoro-2-deoxy-D-glucose), and their diagnostic and/ortherapeutic use in brain disorders associated with aging, such as,senile dementia and Alzheimer's disease. In certain embodiments, theinositol improves the selectivity and/or uptake of the glucose.

Other aspects, features, objects, and advantages of the presentinvention are apparent in the detailed description that follows. Itshould be understood, however, that the detailed description is given byway of illustration only, not limitation. Various changes andmodifications within the scope of the invention will become apparent tothose skilled in the art from the detailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts a table showing the biodistribution in rats following theinjection of [F-18]-N-2-fluoroethyl-2,8-diazaspiro[4,5]decane-1,3-dioneat three different time points.

FIG. 2 depicts a table showing the biodistribution in rats following theinjection of [F-18]-N-2-fluoropropyl-2,8-diazaspiro[4,5]decane-1,3-dioneat two different time points.

FIG. 3 depicts one approach to the synthesis of two fluorinated2,8-diazaspiro[4,5]decane-1,3-dione derivatives.

FIG. 4 depicts one approach to the synthesis ofN-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione.

FIG. 5 depicts three examples of combinations of2,8-diazaspiro[4,5]decane-1,3-dione and inositol structures.

FIG. 6 depicts one approach to the synthesis of6-O—[N—(N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbonyl]-α,β-D-glucopyranose.

FIG. 7 depicts one approach to the synthesis of1,2,3,4,5-penta-O-benzyl-scyllo-inositol.

FIG. 8 depicts one approach to the synthesis of1-deoxy-1-F-18-fluoro-scyllo-inositol

FIG. 9 depicts one approach to the synthesis of6-O-[(β-D-glucopyranose-6-yl)-succinyl]-scyllo-inositol.

FIG. 10 depicts a table showing the some of the effects of amyloidbuildup.

DETAILED DESCRIPTION

One aspect of the present invention provides embodiments of compounds,compositions and methods for effective administration to a subjectsuffering from amyloidosis-associated pathological conditions, such asAlzheimer's disease (AD). In certain embodiments of the invention, thecompounds are reversible choline esterase inhibitors that show highbrain uptake. In certain embodiments, the compounds inhibit β-(Aβ-)protein folding that causes amyloid plaque formation. Further, incertain embodiments, the compounds are labeled with a PET or SPECTradionuclide, and can be used for diagnosing amyloid deposits inpatients suffering from Alzheimer's disease and/or pathologicalconditions characterized by the presence of amyloid deposits. In certainembodiments, the novel imaging compounds act on both potential Alzheimerdisease progressions.

One aspect of the invention relates to the preparation of fluorinated orradiofluorinated 2,8-diazaspiro[4,5]decane-1,3-diones, and theirdiagnostic and/or therapeutic use in amyloidosis-associated pathologicalconditions. For example in brain disorders associated with aging, suchas, senile dementia and Alzheimer's disease. In certain embodiments, thecompositions comprise an effective amount of a fluorinated2,8-diazaspiro[4,5]decane-1,3-dione, in combination with apharmaceutical carrier in an appropriate dosage. In certain embodiments,the compositions comprise a diagnostic radioimaging amount of a¹⁸F-labeled fluorinated 2,8-diazaspiro[4,5]decane-1,3-dione, incombination with a pharmaceutical carrier in an appropriate dosage.

Another aspect of the invention relates to compounds which are acombination of a scyllo-inositol and a2,8-diazaspiro[4,5]decane-1,3-dione (e.g., wherein a scyllo-inositol iscovalently linked by a cross-linker to a2,8-diazaspiro[4,5]decane-1,3-dione). Some of such compounds may bereversible acetyl choline esterase inhibitors and/or A β-amyloid plaqueformation inhibitors; in addition, as such, some compounds may be usefulfor imaging and treating brain disorders associated with aging, such as,for example, senile dementia and Alzheimer's disease. In certainembodiments, certain compounds which are combinations of ascyllo-inositol and a 2,8-diazaspiro[4,5]decane-1,3-dione may showimproved brain uptake and/or brain bioavailability compared to thescyllo-inositol and/or the 2,8-diazaspiro[4,5]decane-1,3-dione.Therefore, such compounds may improve the effective action of thecompound comprising the combination over one or both of the individualcompounds. In certain embodiments, certain compositions of the inventioncomprise a therapeutically and/or diagnostically effective amount of acomposition comprising a compound which is a combination ofscyllo-inositol and a radiofluorinated2,8-diazaspiro[4,5]decane-1,3-dione, and a pharmaceutical carrier in anappropriate dosage.

Another aspect of the invention relates to combinations of2,8-diazaspiro[4,5]decane-1,3-diones and D-glucose for therapy; andcombinations with [F-18]-2-fluoro-2-deoxy-D-glucose for diagnosis. Incertain embodiments, compositions of the invention comprise atherapeutically and/or diagnostically effective amount of a compositioncomprising a compound which is a combination of D-glucose or[F-18]-2-fluoro-2-deoxy-D-glucose and a2,8-diazaspiro[4,5]decane-1,3-dione, and a pharmaceutical carrier in anappropriate dosage.

Another aspect of the invention relates to the combination ofscyllo-inositol with D-glucose for improving uptake and therapy; andcombinations with [F-18]-2-fluoro-2-deoxy-D-glucose for diagnosis. Incertain embodiments, compositions of the invention comprise atherapeutically and/or diagnostically effective amount of a compositioncomprising a compound which is a combination of D-glucose or[F-18]-2-fluoro-2-deoxy-D-glucose and a scyllo-inositol, and apharmaceutical carrier, in an appropriate dosage.

In certain embodiments, imaging and/or therapeutic agents of the instantinvention may be administered to a subject for identification of amyloiddeposits. A specific imaging method detects amyloid deposits byadministering the imaging agent to the subject and detecting the spatialdistribution of the agent. Differential accumulation of the agent isindicative of AD or an amyloidosis-associated pathological condition andcan be monitored by using a PET or SPECT camera.

DEFINITIONS

Herein a number of terms are used extensively. The following definitionsare provided to facilitate understanding of the invention.

The indefinite articles “a” and “an,” as used herein in thespecification and in the claims, unless clearly indicated to thecontrary, should be understood to mean “at least one.”

The phrase “and/or,” as used herein in the specification and in theclaims, should be understood to mean “either or both” of the elements soconjoined, i.e., elements that are conjunctively present in some casesand disjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e., “one or more” ofthe elements so conjoined. Other elements may optionally be presentother than the elements specifically identified by the “and/or” clause,whether related or unrelated to those elements specifically identified.Thus, as a non-limiting example, a reference to “A and/or B”, when usedin conjunction with open-ended language such as “comprising” can refer,in one embodiment, to A only (optionally including elements other thanB); in another embodiment, to B only (optionally including elementsother than A); in yet another embodiment, to both A and B (optionallyincluding other elements); etc.

As used herein in the specification and in the claims, “or” should beunderstood to have the same meaning as “and/or” as defined above. Forexample, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items. Only terms clearly indicated tothe contrary, such as “only one of” or “exactly one of,” or, when usedin the claims, “consisting of,” will refer to the inclusion of exactlyone element of a number or list of elements. In general, the term “or”as used herein shall only be interpreted as indicating exclusivealternatives (i.e., “one or the other but not both”) when preceded byterms of exclusivity, such as “either,” “one of,” “only one of,” or“exactly one of” “Consisting essentially of,” when used in the claims,shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase “atleast one,” in reference to a list of one or more elements, should beunderstood to mean at least one element selected from any one or more ofthe elements in the list of elements, but not necessarily including atleast one of each and every element specifically listed within the listof elements and not excluding any combinations of elements in the listof elements. This definition also allows that elements may optionally bepresent other than the elements specifically identified within the listof elements to which the phrase “at least one” refers, whether relatedor unrelated to those elements specifically identified. Thus, as anon-limiting example, “at least one of A and B” (or, equivalently, “atleast one of A or B,” or, equivalently “at least one of A and/or B”) canrefer, in one embodiment, to at least one, optionally including morethan one, A, with no B present (and optionally including elements otherthan B); in another embodiment, to at least one, optionally includingmore than one, B, with no A present (and optionally including elementsother than A); in yet another embodiment, to at least one, optionallyincluding more than one, A, and at least one, optionally including morethan one, B (and optionally including other elements); etc.

It should also be understood that, unless clearly indicated to thecontrary, in any methods claimed herein that include more than one stepor act, the order of the steps or acts of the method is not necessarilylimited to the order in which the steps or acts of the method arerecited.

In the claims, as well as in the specification above, all transitionalphrases such as “comprising,” “including,” “carrying,” “having,”“containing,” “involving,” “holding,” “composed of,” and the like are tobe understood to be open-ended, i.e., to mean including but not limitedto. Only the transitional phrases “consisting of” and “consistingessentially of” shall be closed or semi-closed transitional phrases,respectively, as set forth in the United States Patent Office Manual ofPatent Examining Procedures, Section 2111.03.

Amyloidosis is a condition characterized by the accumulation of variousinsoluble, fibrillar proteins in the tissues of a patient. An amyloiddeposit is formed by the aggregation of amyloid proteins, followed bythe further combination of aggregates and/or amyloid proteins.

As used herein, “amyloidosis-associated pathological conditions” refersto a group of disorders caused by abnormal folding of proteins leadingto fibril formation in one or more body organs, systems or soft tissues.These clumps of protein are called amyloid deposits and the accumulationof amyloid deposits causes the progressive malfunction and eventualfailure of the affected organ. Normally, proteins are broken down atabout the same rate as they are produced, but these unusually stableamyloid deposits are deposited more rapidly than they can be brokendown. The accumulation may be localized in one organ or may be systemicsuch that several organs are affected.

Amyloidosis causes few or no symptoms in some people, while producingsevere symptoms and fatal complications in other people. The severity ofthe disease depends on which organs are affected by amyloid deposits.Amyloidosis is twice as common in men as in women and is more commonamong older people.

Many forms of amyloidosis exist, and the disease can be classified intofour groups: primary amyloidosis, secondary amyloidosis, hereditaryamyloidosis, and amyloidosis associated with normal aging.

Primary amyloidosis (light chain amyloidosis) occurs with abnormalitiesof plasma cells, and some people with primary amyloidosis also havemultiple myeloma (cancer of the plasma cells). Typical sites of amyloidbuildup in primary amyloidosis are the heart, lungs, skin, tongue,thyroid gland, intestines, liver, kidneys, and blood vessels.

Secondary amyloidosis may develop in response to various diseases thatcause persistent infection or inflammation, such as tuberculosis,rheumatoid arthritis, and familial Mediterranean fever. Typical sites ofamyloid buildup in secondary amyloidosis are the spleen, liver, kidneys,adrenal glands, and lymph nodes.

Hereditary amyloidosis has been noted in some families, particularlythose from Portugal, Sweden, and Japan. The amyloid-producing defectoccurs because of mutations in specific proteins in the blood. Typicalsites for amyloid buildup in hereditary amyloidosis are the nerves,heart, blood vessels, and kidneys.

Amyloidosis associated with normal aging usually affects the heart. Whatcauses amyloid to build up in the heart, other than age, usually is notknown. Amyloid also accumulates in the brain of people with Alzheimer'sdisease and is thought to play a role in causing Alzheimer's. See FIG.10 for a table showing the effects of amyloid buildup.

Besides its presence in Alzheimer's disease, amyloid deposits has alsobeen shown in diseases such as Mediterranean fever, Muckle-Wellssyndrome, idiopathetic myeloma, amyloid polyneuropathy, amyloidcardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy,hereditary cerebral hemorrhage with amyloidosis, Down's syndrome,Scrapie, Creutzfeldt-Jacob disease, Kum, Gerstamnn-Straussler-Scheinkersyndrome, medullary carcinoma of the thyroid, Isolated atrial amyloid,β₂-microglobulin amyloid in dialysis patients, inclusion body myositis,β₂-amyloid deposits in muscle wasting disease, and Islets of Langerhansdiabetes Type I1 insulinoma.

The term “heteroatom” is art-recognized and refers to an atom of anyelement other than carbon or hydrogen. Illustrative heteroatoms includeboron, nitrogen, oxygen, phosphorus, sulfur and selenium.

The term “alkyl” is art-recognized, and includes saturated aliphaticgroups, including straight-chain alkyl groups, branched-chain alkylgroups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkylgroups, and cycloalkyl substituted alkyl groups. In certain embodiments,a straight chain or branched chain alkyl has about 80 or fewer carbonatoms in its backbone (e.g., C₁-C₈₀ for straight chain, C₃-C₈₀ forbranched chain), and alternatively, about 30 or fewer. Likewise,cycloalkyls have from about 3 to about 10 carbon atoms in their ringstructure, and alternatively about 5, 6 or 7 carbons in the ringstructure. As used herein, “fluoroalkyl” denotes an alkyl where one ormore hydrogens have been replaced with fluorines; “perfluoroalkyl”denotes an alkyl where all the hydrogens have been replaced withfluorines.

Unless the number of carbons is otherwise specified, “lower alkyl”refers to an alkyl group, as defined above, but having from one to aboutten carbons, alternatively from one to about six carbon atoms in itsbackbone structure. Likewise, “lower alkenyl” and “lower alkynyl” havesimilar chain lengths.

The term “alkylene,” is art-recognized, and as used herein, pertains toa bidentate moiety obtained by removing two hydrogen atoms, either bothfrom the same carbon atom, or one from each of two different carbonatoms, of a hydrocarbon compound, which may be aliphatic or alicyclic,or a combination thereof, and which may be saturated, partiallyunsaturated, or fully unsaturated. Examples of linear saturatedC₁₋₁₀alkylene groups include, but are not limited to, —(CH₂)_(n)— wheren is an integer from 1 to 10, for example, —CH₂— (methylene), —CH₂CH₂—(ethylene), —CH₂CH₂CH₂— (propylene), —CH₂CH₂CH₂CH₂— (butylene),—CH₂CH₂CH₂CH₂CH₂— (pentylene) and —CH₂CH₂CH₂CH₂CH₂CH₂— (hexylene).Examples of branched saturated C₁₋₁₀alkylene groups include, but are notlimited to, —CH(CH₃)—, —CH(CH₃)CH₂—, —CH(CH₃)CH₂CH₂—,—CH(CH₃)CH₂CH₂CH₂—, —CH₂CH(CH₃)CH₂—, —CH₂CH(CH₃)CH₂CH₂—, —CH(CH₂CH₃)—,—CH(CH₂CH₃)CH₂—, and —CH₂CH(CH₂CH₃)CH₂—. Examples of linear partiallyunsaturated C₁₋₁₀alkylene groups include, but are not limited to,—CH═CH— (vinylene), —CH═CH—CH₂—, —CH═CH—CH₂—CH₂—, —CH═CH—CH₂—CH₂—CH₂—,—CH═CH—CH═CH—, —CH═CH—CH═CH—CH₂—, —CH═CH—CH═CH—CH₂—CH₂—,—CH═CH—CH₂—CH═CH—, and —CH═CH—CH₂—CH₂—CH═CH—. Examples of branchedpartially unsaturated C₁₋₁₀alkylene groups include, but are not limitedto, —C(CH₃)═CH—, —C(CH₃)═CH—CH₂—, and —CH═CH—CH(CH₃)—. Examples ofalicyclic saturated C₁₋₁₀alkylene groups include, but are not limitedto, cyclopentylene (e.g., cyclopent-1,3-ylene), and cyclohexylene (e.g.,cyclohex-1,4-ylene). Examples of alicyclic partially unsaturatedC₁₋₁₀alkylene groups include, but are not limited to, cyclopentenylene(e.g., 4-cyclopenten-1,3-ylene), and cyclohexenylene (e.g.,2-cyclohexen-1,4-ylene, 3-cyclohexen-1,2-ylene, and2,5-cyclohexadien-1,4-ylene).

The term “aralkyl” is art-recognized and refers to an alkyl groupsubstituted with an aryl group (e.g., an aromatic or heteroaromaticgroup).

The terms “alkenyl” and “alkynyl” are art-recognized and refer tounsaturated aliphatic groups analogous in length and possiblesubstitution to the alkyls described above, but that contain at leastone double or triple bond respectively.

The term “aryl” is art-recognized and refers to 5-, 6- and 7-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, naphthalene, anthracene, pyrene,pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole,pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.Those aryl groups having heteroatoms in the ring structure may also bereferred to as “aryl heterocycles” or “heteroaromatics.” The aromaticring may be substituted at one or more ring positions with suchsubstituents as described herein, for example, halogen, azide, alkyl,aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro,sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl,silyl, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester,heterocyclyl, aromatic or heteroaromatic moieties, trifluoromethyl,cyano, or the like. The term “aryl” also includes polycyclic ringsystems having two or more cyclic rings in which two or more carbons arecommon to two adjoining rings (the rings are “fused rings”) wherein atleast one of the rings is aromatic, e.g., the other cyclic rings may becycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls.

The terms ortho, meta and para are art-recognized and refer to 1,2-,1,3- and 1,4-disubstituted benzenes, respectively. For example, thenames 1,2-dimethylbenzene and ortho-dimethylbenzene are synonymous.

The terms “heterocyclyl”, “heteroaryl”, or “heterocyclic group” areart-recognized and refer to 3- to about 10-membered ring structures,alternatively 3- to about 7-membered rings, whose ring structuresinclude one to four heteroatoms. Heterocycles may also be polycycles.Heterocyclyl groups include, for example, thiophene, thianthrene, furan,pyran, isobenzofuran, chromene, xanthene, phenoxanthene, pyrrole,imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, pyrimidine, phenanthroline, phenazine,phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane,thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactamssuch as azetidinones and pyrrolidinones, sultams, sultones, and thelike. The heterocyclic ring may be substituted at one or more positionswith such substituents as described above, as for example, halogen,alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro,sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl,silyl, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, anaromatic or heteroaromatic moiety, trifluoromethyl, cyano, or the like.

The term “nitro” is art-recognized and refers to —NO₂; the term “halo”or “halogen” is art-recognized and refers to —F, —Cl, —Br or —I, and asused herein also refers to radioactive forms thereof, such as ¹⁸F, ⁷⁶Br,⁷⁷Br, ¹²³I, ¹²⁴I and ¹²⁵I; the term “sulfhydryl” is art-recognized andrefers to —SH; the term “hydroxyl” means —OH; and the term “sulfonyl” isart-recognized and refers to —SO₂ ⁻.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines, e.g., a moiety that may berepresented by the general formulas:

wherein R50, R51, R52 and R53 each independently represent a hydrogen,an alkyl, an alkenyl, —(CH₂)_(m)—R61, or R50 and R51 or R52, takentogether with the N atom to which they are attached complete aheterocycle having from 4 to 8 atoms in the ring structure; R61represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle or apolycycle; and m is zero or an integer in the range of 1 to 8. In otherembodiments, R50 and R51 (and optionally R52) each independentlyrepresent a hydrogen, an alkyl, an alkenyl, or —(CH₂)_(m)—R61. Thus, theterm “alkylamine” includes an amine group, as defined above, having asubstituted or unsubstituted alkyl attached thereto, i.e., at least oneof R50 and R51 is an alkyl group.

The term “acylamino” is art-recognized and refers to a moiety that maybe represented by the general formula:

wherein R50 is as defined above, and R54 represents a hydrogen, analkyl, an alkenyl or —(CH₂)_(m)—R61, where m and R61 are as definedabove.

The term “amido” is art recognized as an amino-substituted carbonyl andincludes a moiety that may be represented by the general formula:

wherein R50 and R51 are as defined above. Certain embodiments of theamide in the present invention will not include imides which may beunstable.

The term “alkylthio” refers to an alkyl group, as defined above, havinga sulfur radical attached thereto. In certain embodiments, the“alkylthio” moiety is represented by one of —S-alkyl, —S-alkenyl,—S-alkynyl, and —S—(CH₂)_(m)—R61, wherein m and R61 are defined above.Representative alkylthio groups include methylthio, ethyl thio, and thelike.

The term “carboxyl” is art recognized and includes such moieties as maybe represented by the general formulas:

wherein X50 is a bond or represents an oxygen or a sulfur, and R55 andR56 represents a hydrogen, an alkyl, an alkenyl, —(CH₂)_(m)—R61 or apharmaceutically acceptable salt, R56 represents a hydrogen, an alkyl,an alkenyl or —(CH₂)_(m)—R61, where m and R61 are defined above. WhereX50 is an oxygen and R55 or R56 is not hydrogen, the formula representsan “ester”. Where X50 is an oxygen, and R55 is as defined above, themoiety is referred to herein as a carboxyl group, and particularly whenR55 is a hydrogen, the formula represents a “carboxylic acid”. Where X50is an oxygen, and R56 is hydrogen, the formula represents a “formate”.In general, where the oxygen atom of the above formula is replaced bysulfur, the formula represents a “thiolcarbonyl” group. Where X50 is asulfur and R55 or R56 is not hydrogen, the formula represents a“thiolester.” Where X50 is a sulfur and R55 is hydrogen, the formularepresents a “thiolcarboxylic acid.” Where X50 is a sulfur and R56 ishydrogen, the formula represents a “thiolformate.” On the other hand,where X50 is a bond, and R55 is not hydrogen, the above formularepresents a “ketone” group. Where X50 is a bond, and R55 is hydrogen,the above formula represents an “aldehyde” group.

The term “carbamoyl” refers to —O(C═O)NRR′, where R and R′ areindependently H, aliphatic groups, aryl groups or heteroaryl groups.

The term “oxo” refers to a carbonyl oxygen (═O).

The terms “oxime” and “oxime ether” are art-recognized and refer tomoieties that may be represented by the general formula:

wherein R75 is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl,aralkyl, or —(CH₂)_(m)—R61. The moiety is an “oxime” when R is H; and itis an “oxime ether” when R is alkyl, cycloalkyl, alkenyl, alkynyl, aryl,aralkyl, or —(CH₂)_(m)—R61.

The terms “alkoxyl” or “alkoxy” are art-recognized and refer to an alkylgroup, as defined above, having an oxygen radical attached thereto.Representative alkoxyl groups include methoxy, ethoxy, propyloxy,tert-butoxy and the like. An “ether” is two hydrocarbons covalentlylinked by an oxygen. Accordingly, the substituent of an alkyl thatrenders that alkyl an ether is or resembles an alkoxyl, such as may berepresented by one of —O-alkyl, —O-alkenyl, —O-alkynyl,—O—(CH₂)_(m)—R61, where m and R61 are described above.

The term “sulfonate” is art recognized and refers to a moiety that maybe represented by the general formula:

in which R57 is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

The term “sulfate” is art recognized and includes a moiety that may berepresented by the general formula:

in which R57 is as defined above.

The term “sulfonamido” is art recognized and includes a moiety that maybe represented by the general formula:

in which R50 and R56 are as defined above.

The term “sulfamoyl” is art-recognized and refers to a moiety that maybe represented by the general formula:

in which R50 and R51 are as defined above.

The term “sulfonyl” is art-recognized and refers to a moiety that may berepresented by the general formula:

in which R58 is one of the following: hydrogen, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl or heteroaryl.

The term “sulfoxido” is art-recognized and refers to a moiety that maybe represented by the general formula:

in which R58 is defined above.

The term “phosphoryl” is art-recognized and may in general berepresented by the formula:

wherein Q50 represents S or O, and R59 represents hydrogen, a loweralkyl or an aryl. When used to substitute, e.g., an alkyl, thephosphoryl group of the phosphorylalkyl may be represented by thegeneral formulas:

wherein Q50 and R59, each independently, are defined above, and Q51represents O, S or N. When Q50 is S, the phosphoryl moiety is a“phosphorothioate”.

The term “phosphoramidite” is art-recognized and may be represented inthe general formulas:

wherein Q51, R50, R51 and R59 are as defined above.

The term “phosphonamidite” is art-recognized and may be represented inthe general formulas:

wherein Q51, R50, R51 and R59 are as defined above, and R60 represents alower alkyl or an aryl.

Analogous substitutions may be made to alkenyl and alkynyl groups toproduce, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls,amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls,carbonyl-substituted alkenyls or alkynyls.

The term “selenoalkyl” is art-recognized and refers to an alkyl grouphaving a substituted seleno group attached thereto. Exemplary“selenoethers” which may be substituted on the alkyl are selected fromone of —Se-alkyl, —Se-alkenyl, —Se-alkynyl, and —Se—(CH₂)_(m)—R61, m andR61 being defined above.

The terms triflyl, tosyl, mesyl, and nonaflyl are art-recognized andrefer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl,and nonafluorobutanesulfonyl groups, respectively. The terms triflate,tosylate, mesylate, and nonaflate are art-recognized and refer totrifluoromethanesulfonate ester, p-toluenesulfonate ester,methanesulfonate ester, and nonafluorobutanesulfonate ester functionalgroups and molecules that contain said groups, respectively.

The definition of each expression, e.g., alkyl, m, n, and the like, whenit occurs more than once in any structure, is intended to be independentof its definition elsewhere in the same structure.

The abbreviations Me, Et, Ph, Tf, Nf, Ts, and Ms represent methyl,ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl,p-toluenesulfonyl and methanesulfonyl, respectively. A morecomprehensive list of the abbreviations utilized by organic chemists ofordinary skill in the art appears in the first issue of each volume ofthe Journal of Organic Chemistry; this list is typically presented in atable entitled Standard List of Abbreviations.

Certain compounds contained in compositions of the present invention mayexist in particular geometric or stereoisomeric forms. In addition,polymers of the present invention may also be optically active. Thepresent invention contemplates all such compounds, including cis- andtrans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers,(L)-isomers, the racemic mixtures thereof, and other mixtures thereof,as falling within the scope of the invention. Additional asymmetriccarbon atoms may be present in a substituent such as an alkyl group. Allsuch isomers, as well as mixtures thereof, are intended to be includedin this invention.

If, for instance, a particular enantiomer of compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, or other reaction.

The term “substituted” is also contemplated to include all permissiblesubstituents of organic compounds. In a broad aspect, the permissiblesubstituents include acyclic and cyclic, branched and unbranched,carbocyclic and heterocyclic, aromatic and nonaromatic substituents oforganic compounds. Illustrative substituents include, for example, thosedescribed herein above. The permissible substituents may be one or moreand the same or different for appropriate organic compounds. Forpurposes of this invention, the heteroatoms such as nitrogen may havehydrogen substituents and/or any permissible substituents of organiccompounds described herein which satisfy the valences of theheteroatoms. This invention is not intended to be limited in any mannerby the permissible substituents of organic compounds.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,“Handbook of Chemistry and Physics”, 67th Ed., 1986-87, inside cover.

As used herein, the term “subject” or “individual” refers to a human orother vertebrate animal. It is intended that the term encompass“patients.”

As used herein, a “combination” of two compounds indicates one compoundwherein the two compounds have been linked by a cross linker. Forexample, examples of combinations between2,8-diazaspiro[4,5]decane-1,3-diones and inositol are shown in FIG. 5.

The term “diagnosis” as used herein refers to methods by which theskilled artisan can estimate and/or determine whether or not a patientis suffering from a given disease or condition. The skilled artisanoften makes a diagnosis on the basis of one or more diagnosticindicators, i.e., a marker, the presence, absence, amount, or change inamount of which is indicative of the presence, severity, or absence ofthe condition.

The kit may also include at least one chelating structure and/or anauxiliary molecule such as mannitol, gluconate, glucoheptonate andtartrate and a tin containing reducing agent.

The term “conjugated” refers to ionically or covalently attached (e.g.,via a crosslinking agent).

A “chelating structure” refers to any molecule or complex of moleculesthat bind to a metal as well as the structure bound to a metal. In someembodiments, the metal can be radioactive (such as ^(99m)Tc, ⁶⁸Cu, ⁶⁴Cuand ⁶⁸Ga). Examples of chelating structures include N₂S₂ structure, aHYNIC (hydrazinonicotinic acid) group-containing structure, a2-methylthiolnicotinic acid group containing structure, a carboxylategroup-containing structure and the like. Additional discussion ofchelating structures is below.

A “radioimaging agent” refers to a composition capable of generating adetectable image upon binding with a target and shall includeradionuclides such as ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I, ^(99m)Tc, ⁶⁸Cu,⁶⁴Cu and ⁶⁸Ga.

A “fluorescence imaging agent” refers to a composition capable ofgenerating a detectable optical imaging upon binding with a target withor without specific wave length of light activation and shall includefluorophores. The preferred fluorescence agents are near infra red lightabsorbing agents.

A “target” refers to an in vivo site to which imaging compounds binds. Apreferred target is a brain tissue from a subject suffering from AD oran amyloidosis-associated pathological condition.

A “targeting molecule” is any molecule or biological entity thatspecifically accumulates in brain tissue from a subject suffering fromAD or an amyloidosis-associated pathological condition.

Radioimaging methods that may be employed in accordance with the presentinventions are known in the art. See U.S. Pat. No. 6,187,286 and U.S.Patent Publication No. 2006/0140859; both of which are herebyincorporated by reference.

In accordance with the invention, the targeting molecule is inassociation with (spatial proximity to) the radionuclide. Spatialproximity between the targeting molecule and the radionuclide may beeffected in any manner which preserves the specificity of the targetingmolecule for its target tissue. For example, spatial proximity betweenthe radionuclide and the targeting molecule may be effected by acovalent or non-covalent chemical bond. Such a chemical bond may beeffected through a chelating substance and/or an auxiliary molecule suchas mannitol, gluconate, glucoheptonate, tartrate, and the like.

Alternatively, spatial proximity between the nuclide and the targetingmolecule may be effected by incorporating the radionuclide and thetargeting molecule in a micelle or liposome, in such a way that theaffinity of the targeting molecule for its target tissue is maintained.Spatial proximity between the radionuclide and the targeting moleculemay also be effected by attaching the radionuclide and the targetingmolecule to a matrix such, as a microsphere or liposomes.

A radionuclide may be incorporated into the imaging agent by covalentbonding directly to an atom of the targeting molecule, or theradionuclide may be noncovalently or covalently associated with thetargeting molecule through a chelating structure or through an auxiliarymolecule such as mannitol, gluconate, glucoheptonate, tartrate, and thelike. When a chelating structure is used to provide spatial proximitybetween the radionuclide and the targeting molecule, the chelatingstructure may be directly associated with the targeting molecule or itmay be associated with the targeting molecule through an auxiliarymolecule such as mannitol, gluconate, glucoheptonate, tartrate, and thelike.

Any suitable chelating structure may be used to provide spatialproximity between the radionuclide and the targeting molecule of theagent through covalent or noncovalent association. Many such chelatingstructures are known in the art. Preferably, the chelating structure isan N₂S₂ structure, an NS₃ structure, an N₄ structure, anisonitrile-containing structure, a hydrazine containing structure, aHYNIC (hydrazinonicotinic acid) group-containing structure, a2-methylthiolnicotinic acid group-containing structure, a carboxylategroup containing structure, and the like. In some cases, chelation canbe achieved without including a separate chelating structure, becausethe radionuclide chelates directly to atom(s) in the targeting moiety,for example to oxygen atoms in the phosphate group(s) or in carboxylategroup(s).

The chelating structure, auxiliary molecule, or radionuclide may beplaced in spatial proximity to any position of the targeting moleculewhich does not interfere with the interaction of the targeting moleculewith its receptor in tumors. The chelating structure, auxiliarymolecule, or radionuclide may be covalently or non-covalently associatedwith any moiety of the targeting molecule except the receptor-bindingmoiety. For example, the chelating structure, auxiliary molecule, orradionuclide may be associated with the phosphate moiety of thetargeting molecule, with the —X— moiety of the targeting molecule.

After the labeling reaction is complete, the reaction mixture mayoptionally be purified using one or more high performance liquidchromatography (HPLC) steps. Any suitable HPLC system may be used if apurification step is performed, and the yield of imaging agent obtainedfrom the HPLC step may be optimized by varying the parameters of theHPLC system, as is known in the art. Any HPLC parameter may be varied tooptimize the yield of the imaging agent of the invention. For example,the pH may be varied, e.g., raised, to decrease the elution time of thepeak corresponding to the imaging agent of the invention.

The invention as embodied in a kit for radioimaging comprises aradioimaging agent described above, in combination with apharmaceutically acceptable carrier such as human serum albumin. Humanserum albumin for use in the kit of the invention may be made in anyway, for example, through purification of the protein from human serumor through recombinant expression of a vector containing a gene encodinghuman serum albumin. Other substances may also be used as carriers inaccordance with this embodiment of the invention, for example,detergents, dilute alcohols, carbohydrates, auxiliary molecules, and thelike. The kit of the invention may of course also contain such otheritems as may facilitate its use, such as syringes, instructions,reaction vials, and the like.

In one embodiment, a kit according to the invention contains from about1 to about 30 mCi of the radionuclide-labeled amyloid imaging agentdescribed herein, in combination with a pharmaceutically-acceptablecarrier. The amyloid imaging agent and carrier may be provided insolution or in lyophilized form. When the amyloid imaging agent andcarrier of the kit are in lyophilized form, the kit may optionallycontain a sterile and physiologically acceptable reconstitution mediumsuch as water, saline, buffered saline, and the like.

The radioimaging agents of the invention may be used in accordance withthe methods of the invention by those of skill in the art, e.g., byspecialists in nuclear medicine, to image tissue in a mammal. Anymammalian tumor may be imaged the imaging agents of the invention.Images are generated by virtue of differences in the spatialdistribution of the imaging agents which accumulate in the varioustissues and organs of the mammal. The spatial distribution of theimaging agent accumulated in a mammal, in an organ, or in a tissue maybe measured using any suitable means, for example, a PET or singlephoton emission computer tomography (SPECT) imaging camera apparatus,and the like.

PET imaging is accomplished with the aid of tracer compounds labeledwith a positron-emitting isotope (Goodman, M. M. Clinical PositronEmission Tomography, Mosby Yearbook, 1992, K. F. Hubner et al., Chapter14). These tracer compounds can be labeled with a positron-emittingradionuclide that includes ¹⁸F and ⁷⁶Br. In general, a PET label, is alabel which is covalently attached to the remainder of a molecule andshould have a half-life of at least about 5-20 minutes, preferably about60 minutes or more. Examples of PET labels include ¹⁸F, ¹³N, ⁷⁶Br(half-life=16.1 hrs), ⁷⁷Br, ¹⁵O, ⁶⁸Ga (half-life=68.3 min), ⁶²Cu(half-life=9.74 min), ⁶⁴Cu (half-life=12.7 hrs), ⁸²Rb (half-life=78sec), and ²⁴I (half-life=4.18 days)

The use of ¹⁸F-labeled compounds in PET has thus far been limited to afew analog compounds. Most notably, 18-fluorodeoxyglucose has beenwidely used in studies of glucose metabolism and localization of glucoseuptake associated with brain activity. More recently, other analogs,such as ¹⁸F-methyl choline (for prostate cancers; see Cancer Res. 2001,6, 110), ¹⁸F-fluorothymidine (for lung tumors; see J. Nucl. Med. 2003,44, 1426; and Eur. J. Nuc. Mol. Imaging. 2003, 30, 1407) andO-(2-[¹⁸F]fluoroethyl)-L-tyrosine (U.S. Pat. No. 7,138,540; herebyincorporated by reference), have also been employed in PET imaging. Forexamples of ¹⁸F-labeling imaging agents see: Eur. J. Med. Chem. 1994,29, 115; Eur. J. Med. Chem. 1994, 29, 955; J. Heterocyclic Chem. 1993,30, 1337; Organic Process Research & Development 2005, 9(6), 774; J.Med. Chem. 2005, 48(16), 5290; J. Med. Chem. 1990, 33, 1482; NuclearMedicine and Biology 2001, 28(6), 683; and Nuclear Medicine and Biology2004, 31(4), 483.

For SPECT imaging, the inventive compound can be labeled with aγ-emitting nuclide, such as, ^(99m)Tc, ¹¹¹In, ⁶⁷Ga, ²⁰¹Tl, ¹²³I, ¹³³Xeand others.

For fluorescence tomography imaging, the inventive compound can beconjugated to a near infra red moiety, such as CY5 (Cyanine dye).Fluorescence tomography is under development.

The imaging agents of the instant invention are used in the followingmanner. An effective amount of an imaging agent comprising at least onetargeting molecule and a nuclide (from 1 to 50 mCi) may be combined witha pharmaceutically-acceptable carrier for use in imaging studies. Inaccordance with the invention, “an effective amount” of the imagingagent of the invention is defined as an amount sufficient to yield anacceptable image using equipment which is available for clinical use. Aneffective amount of the imaging agent of the invention may beadministered in more than one injection. Effective amounts of theimaging agent of the invention will vary according to factors such asthe degree of susceptibility of the individual, the age, sex, and weightof the individual, idiosyncratic responses of the individual, and thedosimetry. Effective amounts of the imaging agent of the invention willalso vary according to instrument and film-related factors. Optimizationof such factors is well within the level of skill in the art.

As used herein, “pharmaceutically-acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic agents, absorption delaying agents, and the like.Pharmaceutically-acceptable carriers are materials, useful for thepurpose of administering the compounds in the method of the presentinvention, which are preferably non-toxic, and may be solid, liquid, orgaseous materials, which are otherwise inert and pharmaceuticallyacceptable, and are compatible with the compounds of the presentinvention. Examples of such carriers include oils such as corn oil,buffers such as PBS, saline, polyethylene glycol, glycerin,polypropylene glycol, dimethylsulfoxide, an amide such asdimethylacetamide, a protein such as albumin, and a detergent such asTween 80, mono- and oligopolysaccharides such as glucose, lactose,cyclodextrins and starch.

The formulation used in the present invention may also containstabilizers, preservatives, buffers, antioxidants, or other additivesknown to those of skill in the art. The use of such media and agents forpharmaceutically-active substances is well known in the art.Supplementary active compounds can also be incorporated into the imagingagent of the invention. The imaging agent of the invention may furtherbe administered to an individual in an appropriate diluent or adjuvant,co-administered with enzyme inhibitors or in an appropriate carrier suchas human serum albumin or liposomes. Pharmaceutically-acceptablediluents include sterile saline and other aqueous buffer solutions.Adjuvants contemplated herein include resorcinols, non-ionic surfactantssuch as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether.Enzyme inhibitors include pancreatic trypsin inhibitor,diethylpyrocarbonate, and trasylol. Liposomes inhibitors includewater-in-oil-in-water CGF emulsions, as well as conventional liposomes(see J. Neuroimmunol. 1984, 7, 27).

As described herein, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically-acceptable salts withpharmaceutically-acceptable acids. The term “pharmaceutically-acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting a purifiedcompound of the invention in its free base form with a suitable organicor inorganic acid, and isolating the salt thus formed during subsequentpurification. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate,valerate, oleate, palmitate, stearate, laurate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,naphthylate, mesylate, glucoheptonate, lactobionate, andlaurylsulphonate salts and the like. See J. Pharm. Sci. 1977, 66, 1-19.

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicylic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically-acceptable salts with pharmaceutically-acceptablebases. The term “pharmaceutically-acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting the purified compoundin its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically-acceptable metal cation,with ammonia, or with a pharmaceutically-acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.(See, for example, J. Pham. Sci. 1977., supra)

Preferably, the imaging agent of the present invention is administeredintravenously, and the imaging agent will be formulated as a sterile,pyrogen-free, parenterally-acceptable aqueous solution. The preparationof such parenterally-acceptable solutions, having due regard to pH,isotonicity, stability, and the like, is within the skill in the art. Apreferred formulation for intravenous injection should contain, inaddition to the imaging agent, an isotonic vehicle such as SodiumChloride Injection, Ringer's Injection, Dextrose Injection, Dextrose andSodium Chloride Injection, Lactated Ringer's Injection, or other vehicleas known in the art.

The amount of imaging agent used for diagnostic purposes and theduration of the imaging study will depend upon the nature and severityof the condition being treated, on the nature of therapeutic treatmentswhich the patient has undergone, and on the idiosyncratic responses ofthe patient. Ultimately, the attending physician will decide the amountof imaging agent to administer to each individual patient and theduration of the imaging study.

The diagnostic imaging amounts are preferably about 3 to 15 millicuries(mCi) for a 70 kg normal adult, more preferably being about 1-25 mCi fora 70 kg normal adult.

The ultimate solution form is preferably sterile. Sterilization can beaccomplished by any art recognized technique, including but not limitedto, addition of antibacterial of antifungal agents, for example,paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

More specifically, the compounds that can be formulated into apharmaceutical composition include a therapeutically-effective amount ofthe compound and a pharmaceutically-acceptable carrier. Thetherapeutically-effective amount of the compound and the specificpharmaceutically-acceptable carrier will vary depending upon, e.g., theage, weight, sex of the subject, the mode of administration, and thetype of viral condition being treated.

In a particular aspect, the pharmaceutical composition which can be usedincludes the compounds of the present invention in effective unit dosageform. As used herein, the term “effective unit dosage” or “effectiveunit dose” is used herein to mean a predetermined amount sufficient tobe effective against AD or the like.

The pharmaceutical compositions may contain the compound used in themethod of this invention in an amount of from 0.01 to 99% by weight ofthe total composition, preferably 0.1 to 80% by weight of the totalcomposition. For oral administration, the compound is generallyadministered in an amount of 0.1 g/body to 15 g/body, preferably 0.5g/body to 5 g/body. For intravenous injection, the dose may be about 0.1to about 30 mg/kg/day, preferably about 0.5 to about 10 mg/kg/day. Ifapplied topically as a liquid, ointment, or cream, the compound may bepresent in an amount of about 0.1 to about 50 mg/mL, preferably about0.5 to 30 mg/mL of the composition. Fluorescence agents will beadministered in several μg/kg to several mg/kg. For example, 1-10 mg/kg.

When the compounds according to the invention are formulated forinjection, the dose may be presented in unit dose form in ampoules or inmulti-dose containers with added pharmaceutically-acceptable adjuvantssuch as a preservative.

In addition, the compositions may take forms such as suspensions,solutions, or emulsions in oily or aqueous vehicles, and may containformulating agents, such as suspending, stabilizing, or dispersingagents, isotonic agents and/or dissolving co-solvents conventionallycited in the pharmaceutical art.

For systemic administration, the daily dosage as employed for adulthuman treatment will range from about 0.1 mg/kg to about 150 mg/kg,preferably about 0.2 mg/kg to about 80 mg/kg.

One aspect of the invention relates to a compound, or a pharmaceuticallyacceptable salt thereof, represented by formula I:

wherein, independently for each occurrence,

X is —O— or —S—;

Y is —O—, —S—, —C(R¹)₂—, —N(R⁵)— or —N[(C═O)R¹]—;

L is —R³, —C(═O)R³, —C(═O)[C(R¹)₂]_(p)R³, —C(═O)[C(R¹)₂]_(p)C(═O)R³,—[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)R³, —[C₁₋₁₀alkylene]R³,—C(═O)[C₁₋₁₀alkylene]R³, —[C₁₋₁₀alkylene]C(═O)R³,—C(═O)[C₁₋₁₀alkylene]C(═O)R³, an unsubstituted alkyl, or alkylsubstituted with one or more substituents selected from the groupconsisting of halo, azido, hydroxy, alkoxy, aryloxy, heteroaryloxy,amino, alkylamino, arylamino, acylamino, heteroarylamino, nitro,sulfhydryl, imino, amido, phosphonate, phosphinate, oxycarbonyl,acyloxy, silyl, thioether, sulfo, sulfonate, sulfonyl, sulfonamido,formyl, cyano, isocyano, —(C(R¹)₂)_(q)C(R¹)₃ or a chelating structure;

R¹ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano,isocyano or a chelating structure;

R² is —R³, —C(═O)R³, —C(═O)[C(R¹)₂]_(p)R³, —C(═O)[C(R¹)₂]_(p)C(═O)R³,—[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)R³, —[C₁₋₁₀alkylene]R³,—C(═O)[C₁₋₁₀alkylene]R³, —[C₁₋₁₀alkylene]C(═O)R³,—C(═O)[C₁₋₁₀alkylene]C(═O)R³, an unsubstituted alkyl, or alkylsubstituted with one or more substituents selected from the groupconsisting of halo, azido, hydroxy, alkoxy, aryloxy, heteroaryloxy,amino, alkylamino, arylamino, acylamino, heteroarylamino, nitro,sulfhydryl, imino, amido, phosphonate, phosphinate, oxycarbonyl,acyloxy, silyl, thioether, sulfo, sulfonate, sulfonyl, sulfonamido,formyl, cyano, isocyano, —(C(R¹)₂)_(q)C(R¹)₃ or a chelating structure;

R³ is hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, aralkyl, heteroaralkyl,

R⁴ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano,isocyano, —OR⁵, —SR⁵, —N(R⁵)₂, —(C(R¹)₂)_(q)C(R¹)₃ or a chelatingstructure;

R⁵ is hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, aralkyl, heteroaralkyl, acyl, —(C(R¹)₂)_(q)C(R¹)₃ or achelating structure;

m is 1, 2 or 3;

n is 1, 2 or 3;

m plus n is 3 or 4

p is 1-10 inclusive; and

q is 0-10 inclusive;

provided that when R² is an unsubstituted alkyl, or alkyl substitutedwith one or more substituents selected from the group consisting ofhalo, azido, hydroxy, alkoxy, aryloxy, heteroaryloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, oxycarbonyl, acyloxy, silyl, thioether, sulfo,sulfonate, sulfonyl, sulfonamido, formyl, cyano, isocyano,—(C(R¹)₂)_(q)C(R¹)₃ or a chelating structure, then L is —R³, —C(═O)R³,—C(═O)[C(R¹)₂]_(p)R³, —C(═O)[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)C(═O)R³,—[C(R¹)₂]_(p)R³, —[C₁₋₁₀alkylene]R³, —C(═O)[C₁₋₁₀alkylene]R³,—[C₁₋₁₀alkylene]C(═O)R³, —C(═O)[C₁₋₁₀alkylene]C(═O)R³; and when L is anunsubstituted alkyl, or alkyl substituted with one or more substituentsselected from the group consisting of halo, azido, hydroxy, alkoxy,aryloxy, heteroaryloxy, amino, alkylamino, arylamino, acylamino,heteroarylamino, nitro, sulfhydryl, imino, amido, phosphonate,phosphinate, oxycarbonyl, acyloxy, silyl, thioether, sulfo, sulfonate,sulfonyl, sulfonamido, formyl, cyano, isocyano, —(C(R¹)₂)_(q)C(R¹)₃ or achelating structure, then R² is —R³, —C(═O)R³, —C(═O)[C(R¹)₂]_(p)R³,—C(═O)[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)R³,—[C₁₋₁₀alkylene]R³, —C(═O)[C₁₋₁₀alkylene]R³, —[C₁₋₁₀ alkylene]C(═O)R³,—C(═O)[C₁₋₁₀alkylene]C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises at least one ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I or ¹²⁵I.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises at least one ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises only one chelating structure.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises only one chelating structure; and ^(99m)Tc, ⁶⁸Cu,⁶⁴Cu or ⁶⁸Ga chelated to the chelating structure.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein Y is—C(R¹)₂—.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein Y is —CH₂—.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein Y is —O—.For examples of 4-oxa-analogs of the muscarinic agonist2-ethyl-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione, see J. Med. Chem.1993, 36, 2292, which is hereby incorporated by reference.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R¹ ishydrogen.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein m is 1; andn is 2.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein m is 1; andn is 3.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein m is 2; andn is 2.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH—; R¹ is hydrogen; m is 2; and n is 2.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² isunsubstituted alkyl.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² is —CH₃,—CH₂CH₃, —CH₂CH₂CH₃, or —CH(CH₃)CH₃.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² is alkylsubstituted with —F, —Cl, —Br or —I.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² is alkylsubstituted with —F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² is alkylsubstituted with —¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² is—CH₂CH₂ ¹⁸F or —CH₂CH¹⁸FCH₃

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; and R² is alkyl substitutedwith —F, —Cl, —Br or —I.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; and R² is alkyl substitutedwith —F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; and R² is alkyl substitutedwith —¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; and R² is —CH₂CH₂ ¹⁸F or—CH₂CH¹⁸FCH₃

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is R³; andR³ is alkyl.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is R³; andR³ is —CH₃.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—F, —Cl, —Br or —I; L is R³; and R³ is alkyl.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—F; L is R³; and R³ is alkyl.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—¹⁸F; L is R³; and R³ is alkyl.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is —CH₂CH₂ ¹⁸F or—CH₂CH¹⁸FCH₃; L is R³; and R³ is —CH₃.

One aspect of the invention relates to a fluorinated2,8-diazaspiro[4,5]decane-1,3-dione compound, or pharmaceuticallyacceptable salt thereof.

One aspect of the invention relates to a ¹⁸F-fluorinated2,8-diazaspiro[4,5]decane-1,3-dione compound, or a pharmaceuticallyacceptable salt thereof.

One aspect of the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein the compoundis [F-18]-N-2-fluoroethyl-2,8-diazaspiro[4,5]decane-1,3-dione or[F-18]-N-2-fluoroethyl-2,8-diazaspiro[4,5]decane-1,3-dione.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³, —C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or—[CH₂]_(p)C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)CH₂CH₂C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³, —C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or—[CH₂]_(p)C(═O)R³; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³, —C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or—[CH₂]_(p)C(═O)R³; R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)CH₂CH₂C(═O)R³; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—¹⁸F; L is —C(═O)R³, —C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or—[CH₂]_(p)C(═O)R³; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—¹⁸F; L is —C(═O)R³, —C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or—[CH₂]_(p)C(═O)R³; R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is —CH₂CH₂ ¹⁸F or—CH₂CH¹⁸FCH₃; L is —C(═O)R³; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is —CH₂CH₂ ¹⁸F or—CH₂CH¹⁸FCH₃; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —F, —Cl, —Br or —I; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —F; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —¹⁸F; and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is unsubstituted alkyl oralkyl substituted with —F, —Cl, —Br, or —I; L is —C(═O)R³,—C(═O)[CH₂]_(p)R³, —C(═O)[CH₂]_(p)C(═O)R³, or —[CH₂]_(p)C(═O)R³; and R³is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl; L is —C(═O)R³ or—C(═O)[CH₂]_(p)C(═O)R³; R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is unsubstituted alkyl oralkyl substituted with —F, —Cl, —Br, or —I; L is —C(═O)CH₂CH₂C(═O)R³; R³is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—F; L is —C(═O)CH₂CH₂C(═O)R³; R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is alkyl substituted with—¹⁸F; L is —C(═O)CH₂CH₂C(═O)R³; R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is —CH₂CH₂ ¹⁸F or—CH₂CH^(18F)CH₃; L is —C(═O)CH₂CH₂C(═O)R³; R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is; L is —C(═O)R³ or—C(═O)[CH₂]_(p)C(═O)—; R³ is

W is —F; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein X is —O—; Yis —CH₂—; R¹ is hydrogen; m is 2; n is 2; R² is unsubstituted alkyl; Lis —C(═O)R³ or —C(═O)CH₂CH₂C(═O)—; R³ is

W is —¹⁸F; and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein the compoundcomprises a radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R¹ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R¹ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R² comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R³ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R³ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁴ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁴ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

One aspect of the invention relates to a compound, or a pharmaceuticallyacceptable salt thereof, represented by formula II:

wherein, independently for each occurrence,

L is —H, —C(═O)R³, —C(═O)[C(R¹)₂]_(p)R³, —C(═O)[C(R¹)₂]_(p)C(═O)R³,—[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)R³, —[C₁₋₁₀alkylene]R³,—C(═O)[C₁₋₁₀alkylene]R³, —[C₁₋₁₀alkylene]C(═O)R³ or—C(═O)[C₁₋₁₀alkylene]C(═O)R³;

R¹ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano, orisocyano;

R⁴ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano,isocyano, —OR⁵, —SR⁵, —N(R⁵)₂, —(C(R¹)₂)_(q)C(R¹)₃ or a chelatingstructure;

R⁵ is hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, aralkyl, heteroaralkyl, acyl, —(C(R¹)₂)_(q)C(R¹)₃ or achelating structure.

R⁷ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano,isocyano, —OR⁵, —SR⁵, —N(R⁵)₂, —(C(R¹)₂)_(q)C(R¹)₃ or a chelatingstructure;

p is 1-10 inclusive; and

q is 0-10 inclusive;

provided that when L is —H, one R⁷ is fluoro; and the other R⁷ arehydroxy, alkoxy, aryloxy, heteroaryloxy, aralkyloxy, orheteroaralkyloxy.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises at least one ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I or ¹²⁵I.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises at least one ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises only one chelating structure.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein saidcompound comprises only one chelating structure; and ^(99m)Tc, ⁶⁸Cu,⁶⁴Cu or ⁶⁸Ga chelated to the chelating structure.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³—.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)[CH₂]_(p)C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)CH₂CH₂C(═O)R³.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ is —H,—F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ is —H,—F, —Cl, —Br, —I, or —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ is —F or—OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ is —¹⁸For —OH; provided that only one R⁷ is —¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; and R⁷ is —H, —F, —Cl, —Br, —I,or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³; and R⁷ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)[CH₂]_(p)C(═O)R³; and R⁷ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³; and R⁷ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)CH₂CH₂C(═O)R³; and R⁷ is OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³; and R⁷ is R⁷ is —F or —OH; provided that only one R⁷ is —F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)CH₂CH₂C(═O)R³; and R⁷ is —F or —OH; provided that only one R⁷ is—F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³; and R⁷ is R⁷ is —¹⁸F or —OH; provided that only one R⁷ is—¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)CH₂CH₂C(═O)R³; and R⁷ is —¹⁸F or —OH; provided that only one R⁷ is—¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —F, —Cl, —Br or —I; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —F; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, R³ is

W is —¹⁸F; and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; and R³ is

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; and R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; R³ is

and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; R³ is

and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; R³ is

W is —F, —Cl, —Br or —I; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; R³ is

W is —F; and R⁴ is —OR⁵.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is—C(═O)R³ or —C(═O)[C(R¹)₂]_(p)C(═O)R³; R⁷ is —H, —F, —Cl, —Br, —I, or—OR⁵; R³ is

W is —¹⁸F; and R⁴ is —OH.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein L is —H, oneR⁷ is fluoro; and the other R⁷ are hydroxy, alkoxy, aryloxy,heteroaryloxy, aralkyloxy, or heteroaralkyloxy.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein the compoundis 1-deoxy-1-fluoro-scyllo-inostiol or 1-deoxy-1-fluoro-myo-inositol.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein the compoundcomprises a radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R¹ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R¹ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁴ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁴ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ comprisesthe radioimaging agent.

In certain embodiments, the present invention relates to any one of theaforementioned compounds and attendant definitions, wherein R⁷ comprisesthe radioimaging agent; and the radioimaging agent is ¹⁸F.

One aspect of the present invention relates to a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound, or a pharmaceutically acceptable form thereof; wherein thecompound is any of the compound described herein.

In certain embodiments, the present invention relates to any one of theaforementioned pharmaceutical composition and attendant definitions,wherein said compound is a compound of formula I or II and attendantdefinitions thereto.

One aspect of the present invention relates to a method for amyloidimaging a subject suffering from an amyloidosis-associated pathologicalcondition, or treating a subject suffering from amyloidosis-associatedpathological condition, comprising the step of:

administering a compound, or a composition comprising a pharmaceuticallyacceptable carrier and the compound or a pharmaceutically acceptableform thereof, wherein the compound is represented by any of the compounddescribed herein.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said compoundis a compound of formula I or II and attendant definitions thereto.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein saidamyloidosis-associated pathological condition is Alzheimer's disease.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said compoundcomprises at least one ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I or ¹²⁵I.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein the compoundis

One aspect of the present invention relates to a method for treating asubject suffering from an amyloidosis-associated pathological condition,comprising the step of:

administering a compound, or a composition comprising a pharmaceuticallyacceptable carrier and the compound or a pharmaceutically acceptableform thereof, wherein the compound is represented by any of thecompounds described herein.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said compoundis a compound of formula I or II and attendant definitions thereto.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein saidamyloidosis-associated pathological condition is Alzheimer's disease.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said treatmentinhibits the formation of plaque.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said compoundis a combination of RS-86 and inositol, a combination of RS-86 andglucose, a combination of inositol and glucose, or a disaccharidecomprising inositol and glucose.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein saidcombination shows increased uptake over one or both of the constituentsthereof.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein saidcombination shows increased uptake over one or both of the constituentsthereof, in cells of the blood, heart, lung, liver, spleen, kidney,adrenal gland, stomach, gi tract, gonads, skeletal muscle, bone, orbrain.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein saidcombination shows increased uptake over one or both of the constituentsthereof, in the brain.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein said compoundis fluorinated.

In certain embodiments, the present invention relates to any one of theaforementioned methods and attendant definitions, wherein the compoundis

EXEMPLIFICATION

It should be understood that the above-described embodiments and thefollowing examples are given by way of illustration, not limitation.Various changes and modifications within the scope of the presentinvention will become apparent to those skilled in the art from thepresent description.

General Reaction Materials and Methods of Analysis

All reactions were carried out in dry glassware (oven at 150° C. for 12hrs) unless otherwise noted and a steady stream of dry N₂ gas was usedto prevent moisture-air off the reaction system. All chemicals andsolvents for moisture-sensitive reactions were purchased from Aldrichchemical company and used as supplied. Ethyl ether was distilled fromsodium-benzophenone immediately prior to use. Dichloromethane wasdistilled from CaH or P₂O₅ prior to use. DMF was distilled under reducedpressure onto activated 4 angstrom molecular sieves or purchased as drysolvent for immediate use. Syringes possessing Teflon or rubberseals/plungers were dried (24 h) using an evacuated Abderhalden dryingapparatus at 56° C. (acetone) with KOH as the desiccant and the sameAbderhalden drying apparatus at 64° C. (methyl alcohol) with P₂O₅ wasused for sample drying purpose. Reactions carried out at −50° C.employed the frozen slurry of 50% CaCl₂ solution with powdered dry-ice.¹H NMR spectra were recorded on a 300 MHz Varian Mercury instrument andreported in parts per million (ppm), with residual CDCl₃ referenced at7.26 ppm and Me₄Si at 0.00 ppm. Multiplicity, coupling constant (Hz),and proton count follow each peak assignment. Elemental analyses wereperformed by Robertson Laboratories, Madison, N.J., and were within+/−0.4% of theoretical values unless otherwise indicated. Analyticpolyester TLC plates were purchased from Aldrich (Silica gel thickness:250 μm, pore size 60 angstroms, 20×20 cm, Fluorescent indicator, Cat.No. Z122785). TLC plates were visualized using a 254 nm/366 nm UV lampand 5% ethanol solution of phosphomolybdic acid or an iodine/silica gelmixture. Preparative TLC plates were purchased from Sigma-Aldrich(silica gel on glass, 2000 μm, 20×20 cm, fl. Ind. Cat No. Z513040).Chromatographic separations were made using Silia-P Flask silica gel(Silicycle Chemical Division, Quebec, QC, Canada), particle size: 40-63μm, 60 angstroms. HRMS determinations were done by Molecular BiologyCore Facilities, DFCI, MA in the electron-impact (EI) or fast-atombombardment (FAB) mode.

Example 1

FIG. 3 depicts one approach to the synthesis of two novel fluorinated2,8-diazaspiro[4,5]decane-1,3-dione derivatives. Synthetic protocols forthe synthesis of the compounds depicted in FIG. 3 are presented below.

[A] Synthesis of ethyl (1-methyl-4-piperidylidene)cyanoacetate (1; FIG.3). See U.S. Pat. No. 3,056,796, hereby incorporated by reference;Bioorganic & Medicinal Chemistry Letters 2002, 12, 1103; and J. Med.Chem. 2004, 47 (8), 2037. To a stirred solution of 1-methyl-4-piperidone(10.0 g, 88.3 mmol) and ethyl cyanoacetate (13.0 g, 115.2 mmol) inCH₂Cl₂ (100 mL) was added Et₃N (17.9 g, 177.0 mmol). Then, crushed 4angstrom molecular sieves (9.0 g) were added and the mixture was stirredat room temperature overnight. The reaction mixture was filtered throughCelite, and the filtrate was concentrated to give the product as a clearred syrup (15 g; yield 82%). ¹H NMR (CDCl₃-d) δ 1.31 (t, 3H, CH₂CH₃),2.28 (s, 3H, N—CH₃), 2.50 (t, 2H, J_(ab)=5.7 Hz, CH₂N), 2.57 (t, 2H,J_(ab)=6.0 Hz, CH₂N), 2.77 (t, 2H, J_(ab)=6.0 Hz, CH₂C═C), 3.12 (t, 2H,J_(ab)=5.7 Hz, CH₂C═C). It was used directly in the following stepwithout further purification.

[B] Synthesis of 1-methyl-4-cyanomethylpiperidine-4-carbonitrile (2;FIG. 3). The ethyl ester 1 (15 g, 72.1 mmol) obtained from previous stepwas dissolved in ethanol (150 ml). A solution of KCN (28.7 g, 442 mmol)in H₂O (60 mL). The red solution was heated to reflux for 4 h, thensolvent was evaporated under the reduced pressure to a dark-red syrupwhich was extracted with EtOAc (4×100). The combined organic extract waswashed with brine, dried over MgSO₂, and concentrated to yield a redclear oil. Chromatography using 10% methanol in CH₂Cl₂ furnished puredinitrile compound (8.8 g; yield 74.9%). ¹H NMR (CDCl₃-d) δ 1.75 (m, 2H,CH₂N), 2.06 (d, 2H, CH₂N), 2.30 (m, 2H, CH₂C═N), 2.33 (s, 3H, NCH₃),2.73 (s, 2H, CH₂CN), 2.87 (d, 2H, CH2C═N).

[C] Synthesis of 1-methyl-4-carboxymethylpiperidine-4-carboxylic acidhydrochloride (3; FIG. 3). See J. Med. Chem. 2004, 47 (8), 2037. Asuspension of dinitrile 2 (14.4 g, 88.3 mmol) in concentrated HCl (250mL) was refluxed for 36 h. Then the pale yellow solution wasconcentrated under the reduced pressure to dryness to afford the diacid3 as an off-white solid, as its hydrochloride salt (19.1 g; yield 91%).¹H NMR (DMSO-d6) δ 1.92 (t, 2H, CH₂N), 2.17 (t, 2H, CH₂N), 2.46 (s, 1H,CH₂CCO₂H), 2.72 (s, 3H, NCH₃), 2.84 (m, 2H, CH₂CCO₂H), 3.24 (q, 1H,CH₂CCO₂H), 3.33 (s, 2H, CH₂CO₂H), 10.88 (m, 2H, COOH).

[D] Synthesis of 8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione (4; FIG.3). See J. Med. Chem. 1988, 31(8), 1598. The diacid HCl 3 (16.4 g, 69mmol) was dissolved in 30 mL conc. NH₄OH and heated to reflux for 1 hourand then the volatile components of the solution were distilled off atnormal pressure, the residue was heated at 250° C. for 5 h. Aftercooling, the residue was treated with hot water (10 mL), 50% K₂CO₃ (28mL), and using 50% NaOH (1 mL) the pH of the solution was adjusted to12. Then the mixture was extracted with chloroform (3×100 mL) andconcentrated under the reduced pressure to provide a slight brown solid.The crude product was purified by crystallization in 2-propanol to givea yellow crystal (7.6 g; yield 60%). ¹H NMR (CDCl₃-d) δ 1.55 (d, 2H,CH₂N), 2.03 (t, 2H, CH₂C), 2.12 (t, 2H, CH₂N), 2.28 (s, 3H, NCH₃), 2.56(s, 2H, CH₂CO), 2.87 (d, 2H, CH₂C).

Note: For the preparation of compound 4, the product was easy todecompose and had caused lower yield under the condition cited fromliterature, higher temperature 230-280° C. in vacuo. The condition wasmodified by controlling the temperature at 250° C. at the normalpressure to increase the yield from literature 31% to 60%.

[E] Synthesis of2-(2-hydroxyethyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione (5; FIG.3). See J. Med. Chem. 1988, 31(8), 1598. A solution of compound 4 (1.8g, 10 mmol) dissolved in fresh distilled DMF (10 mL) was treated with95% NaH (0.28 g, 11 mmol) in mineral oil and the mixture was stirred for1.5 h at 50° C. and then there was added a solution of 2-bromoethanol(1.58 g, 12 mmol) in DMF (6 mL) over the period of 50 min and themixture was stirred for 4 h at 75 to 80° C. Then the mixture wasfiltered with 1 g active charcoal carbon and the filtrate was evaporatedin vacuo. The residue was extracted with chloroform (3×50 mL) and theprecipitated salts were filtered off, the filtrate was filtered througha layer of neutral Al₂O₃ (activity II). The solvent was evaporated underthe reduced pressure to give a colorless semi-solid. The crude productcan't be perfectly purified by crystallization with a mixture of2-propanol:hexanes (20:5). The pure product was obtained as a colorlesspowder by flash chromatography with 15% methanol in CH₂Cl₂ (1.12 g;yield 50%). ¹H NMR (CDCl₃-d) δ 1.49 (d, 2H, CH₂N), 1.98 (t, 2H, CH₂C),2.08 (t, 2H, CH₂N), 2.25 (s, 3H, NCH₃), 2.51 (s, 2H, CH₂CO), 2.81 (d,2H, CH₂C), 3.63 (t, 2H, NCH₂CH₂), 3.69 (t, 2H, CH₂OH).

Note: In the preparation of spiro-compound 5, using the samepurification method provided in literature was failed to obtain a pureproduct, a need for flash chromatography was required. Elution with 15%methanol in CH₂Cl₂ afforded a 50% yield that was lower than that (76%)reported in literature. The product has a weak UV absorption so its TLCdetection is difficult too. I₂/Silica gel was used for chromatographydetection of the product.

[F] Synthesis of2-(2-methanesulfonylethyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-done(6; FIG. 3). In a 25-ml round-bottom flask was dissolved compound 5 (0.3g, 1.3 mmol) in anhydrous dichloromethane (5 mL). To this solution wasadded triethylamine (0.27 g, 2.6 mmol), the reaction mixture was cooledto 0° C. with an ice-NaCl bath. Methanesulfonyl chloride was addeddropwise, and the reaction mixture was stirred under nitrogen for 2 h,allowing the ice bath to expire and stirred at room temperature for 1 h.Then the reaction mixture was diluted with dichloromethane (20 mL),washed with a sat. solution of NaHCO₃, then brine, dried over MgSO₄, andthe solvent was evaporated under the reduced pressure to provide ayellow clear oil. The product was purified by flash chromatographyeluted with a mixture of 10% methanol in dichloromethane to give acolorless clear syrup which evaporated under high vacuum to obtain acolorless semi-solid (144 mg; yield 36%). ¹H NMR (CDCl₃-d) δ 1.48 (d,2H, CH₂N), 1.93 (t, 2H, CH₂C), 2.05 (t, 2H, CH₂N), 2.23 (s, 3H, NCH₃),2.51 (s, 2H, CH₂CO), 2.78 (d, 2H, CH₂C), 2.94 (s, 3H, CH₃SO₃), 3.76 (t,2H, NCH₂CH₂), 4.34 (t, 2H, CH₂OSO₂CH₃).

[G] Synthesis of2-(2-hydroxypropyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione (7;FIG. 3). To a solution of compound 4 (1.8 g, 10 mmol) in DMF (10 mL) wasslowly added 95% NaH in mineral oil (0.28 g, 11 mmol), which produced alot of bubbles and additional DMF (5 mL) was added. The mixture wasstirred at 65° C. for 1 h. To the mixture a solution of1-bromo-2-propanol (1.2 g, 12 mmol) in DMF (6 mL) was added over 80 minuntil the brown bubbles all dissolved and the mixture became a clearyellow solution, then the temperature of the oil bath was raised up to70° C. and kept overnight. The mixture was added charcoal carbon (0.1 g)and filtered through a layer of Celite. The filtrate was evaporated todryness under the reduced pressure and the residue was extracted withchloroform (3×60 mL). The collected chloroform was filtered off theinsoluble materials and then filtered through a layer of neutral Al₂O₃(activity II). The filtrate was evaporated in the reduced pressure togive a yellow solid. The crude product was purified by crystallizationafter trituration with a mixture of hexanes and 2-propanol (4:1) toprovide a colorless powder (1.3 g; yield 54%). ¹H NMR (CDCl₃-d) δ 1.13(d, 3H, CH₃CH), 1.47 (t, 2H, CH₂N), 1.99 (t, 2H, CH₂N), 2.08 (t, 2H,CH₂C), 2.24 (s, 3H, NCH₃), 2.53 (s, 2H, CH₂CO), 2.82 (t, 2H, CH₂C), 3.48(d, 2H, NCH₂CH), 3.94 (m, 1H, CHOH).

Note: In the preparation of compound 7, the addition of NaH (95%) willcause a large amount of bubbles and heat to be released, so thesuggestion is to use an ice bath at the beginning of the reaction, thentake it away after finishing the NaH addition. After the overnightreaction, charcoal addition is necessary to remove any conjugation ordehydration by-products and Filtration through Al₂O₃ layer is to removepossible acidic by-products. Attempts to react compound 4 with propyleneoxide and sodium hydride in DMF to prepare the product 7 have beenunsuccessful. Although the commercial starting material,1-bromo-2-propanol, contains 30% of its isomer, 2-bromo-1-propanol, itappears that this isomer did not have any negative effect on theseparation of a pure product in the procedure outlined above.

[H] Synthesis of2-(2-methanesulfonylpropyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-done(8; FIG. 3). To a solution of compound 7 (1.3 g, 5.4 mmol) dissolved infreshly distilled CH₂Cl₂ (20 ml) in an ice-salt bath (0° C.) was addedtriethylamine (1.09 g, 6.6 mmol) and then methanesulfonyl chloride wasadded dropwise. The reaction mixture was stirred under nitrogen for 1.5h, allowing the ice-bath to expire, and stirring continued at roomtemperature for 1 h. The mixture was diluted with CH₂Cl₂ (20 mL), andthen washed with sat. NaHCO₃, brine, and dried over MgSO₄. The solutionwas evaporated under the reduced pressure to give a yellow solid. Thecrude product was purified by chromatography eluted with 15% methanol inCH₂Cl₂ to obtain a colorless powder (0.9 g; yield 52%). ¹H NMR (CDCl₃-d)δ 1.40 (d, 3H, CH₃CH), 1.58 (d, 2H, CH₂N), 2.01 (t, 2H, CH₂N), 2.10 (t,2H, CH₂C), 2.28 (s, 3H, NCH₃), 2.53 (s, 2H, CH₂CO), 2.82 (d, 2H, CH₂C),2.94 (s, 3H, CH₃SO₂O), 3.44 (dd, 1H, NCH₂CH), 3.79 (q, 1H, NCH₂CH), 5.04(m, 1H, CHOMs). Elemental Analysis: (C₁₃H₂₂N₂O₅S) Cal: C, 49.04; H,6.96; N, 8.80, S 10.07. Found: C, 48.84; H, 6.75; N, 8.51, S 10.37.

[I] Synthesis of2-(2-fluoroethyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione (9; FIG.3). A solution of compound 4 (0.3 g, 1.65 mmol) dissolved in freshdistilled DMF (2 mL) was treated with 95% NaH (0.05 g, 1.90 mmol) inmineral oil and the mixture was stirred for 1.5 h at 50° C. and thenthere was added a solution of 1-bromo-2-fluoroethane (0.33 g, 2.6 mmol)in DMF (2 mL) over the period of 50 min and the mixture was stirred for4 h at 75 to 80° C. Then the mixture was filtered with active charcoalcarbon (0.01 g) and the filtrate was evaporated in vacuo. The residuewas extracted with chloroform (3×50 mL) and the precipitated salts werefiltered off, the filtrate was filtered through a layer of neutral Al₂O₃(activity II). The solvent was evaporated under the reduced pressure togive a colorless solid. The pure product was obtained as a colorlesspowder by flash chromatography with 15% methanol in CH₂Cl₂ (175 mg;yield 46%). ¹H NMR (CDCl₃-d) δ 1.48 (d, 2H, CH₂N), 1.96 (t, 2H, CH₂C),2.08 (t, 2H, CH₂N), 2.24 (s, 3H, NCH₃), 2.52 (s, 2H, CH₂CO), 2.81 (d,2H, CH₂C), 3.75 (tt, 2H, NCH₂), 4.48 (tt, 2H, CH₂F). Elemental Analysis:(C₁₁H₁₇FN₂O₂) Cal: C, 57.88; H, 7.51; N, 12.27; F 8.32. Found: C, 57.61;H, 7.79; N, 12.13; F 7.73.

Note: In the preparation of compound 9, the method of using compound 6reacted with TBAF (tetrabutylammonium fluoride in THF) in order toreplace the Ms-group with a fluoro-group was unsuccessful due to excessTBAF which could not be separated from the product 9, both have anoverlap R_(f) values in TLC and flash chromatography in elution systemssuch as 10-20% methanol in CH₂Cl₂. See Nucl. Med. Biol. 1993, 20(1), 81.

[J] Synthesis of2-(2-fluoropropyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione (10;FIG. 3). To a solution of compound 4 (1.2 g, 6.5 mmol) in DMF (10 mL)was slowly added 95% NaH in mineral oil (0.2 g, 7.8 mmol). The mixturewas stirred at 65° C. for 1 h. To the mixture, a solution of1-bromo-2-fluoropropane (1.1 g, 7.8 mmol) in DMF (6 mL) was added over80 min until the brown bubbles all dissolved and the mixture became aclear yellow solution, then the temperature of the oil bath was raisedup to 70° C. and kept overnight. After cooling to room temperature, themixture was added charcoal carbon (0.1 g) and filtered through a layerof Celite. The filtrate was evaporated to dryness under the reducedpressure and the residue was extracted with chloroform (3×60 mL). Thecollected chloroform was filtered off the insoluble materials and thenfiltered through a layer of neutral Al₂O₃ (activity II). The filtratewas evaporated in the reduced pressure to give a yellow solid. The crudeproduct was purified by crystallization in hexanes to provide acolorless powder (1.2 g; yield 76%). ¹H NMR (CDCl₃-d) δ 1.33 (q, 3H,CH₃CH), 1.49 (d, 2H, CH₂N), 1.99 (t, 2H, CH₂N), 2.13 (t, 2H, CH₂C), 2.28(s, 3H, NCH₃), 2.57 (s, 2H, CH₂CO), 2.83 (d, 2H, CH₂C), 3.48 (d-q, 1H,NCH₂CHF), 3.78 (d-t, 1H, NCH₂CHF), 4.84 (d-m, 1H, CHF). ElementalAnalysis: (C₁₂H₁₉FN₂O₂) Cal: 59.49; H, 7.90; N, 11.56; F 7.84. Found: C,59.49; H, 8.06; N, 11.55; F 7.69.

Note: In the preparation of compound 10, the method of using compound 7directly reacted with DAST (diethylaminosulfur trifluoride) in CH₂Cl₂has been proved to provide the product 10 only with poor yield 10-20%.See J. Org. Chem. 1975, 40(5), 574.

[K] Synthesis of 1-bromo-2-fluoroethane. See J. Org. Chem. 1975, 40(5),574. 1-Bromoethanol (1.65 g, 12.5 mmol) was added dropwise to a solutionof dimethylaminosulfur trifluoride (DAST) (2.0 g, 12.5 mmol) in diglyme(8 mL) cooled to −50° C. in a bath of dry-ice and acetone. The reactionmixture was warmed to room temperature, and continued to stirring atroom temperature for 1 h. The most volatile portion was distilled at68-75° C. for collection in dry-ice traps by a hood vacuum. Thedistillate was washed with water (4 mL), 5% sodium bicarbonate solution,dried over MgSO₄, and redistilled to give product as a colorless oil(1.4 g; yield 88%). ¹H NMR (CDCl₃-d) δ 3.50 (t-t, 2H, CH₂Br), 4.62 (t-t,2H, CH₂F).

[L] Synthesis of 1-bromo-2-fluoropropane. This compound was synthesizedas reported in the literature; see Organic Syntheses 2004, 10, 128; andOrganic Syntheses 1999, 76, 159. ¹H NMR (CDCl₃-d) δ 1.41 (d-d, 3H,CH₃CH), 3.44 (d-d, 2H, CH₂Br), 4.80 (m-m, 1H, CHF).

Example 2

FIG. 4 depicts one approach to the synthesis ofN-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione (6; FIG. 4). Thepreparation of 2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione 6 was doneaccording to a modified version of a known procedure. See Irie, O.,Iwasaki, G. et al., WO 2004/076455; which is hereby incorporated byreference in its entirety. Synthetic protocols for the synthesis of thecompounds depicted in FIG. 4 are presented below.

[A] Synthesis of 1-benzyl-piperidin-4-ylidene)-cyanoacetate (2; FIG. 4).A solution of 1-benzyl-piperidin-4-one (75.1 g, 0.40 mol) in toluene(9400 mL), ethyl cyanoacetate (50.6 mL, 0.48 mol) and acetic acid (918.2mL, 0.32 mol) was refluxed for 4 hr. The mixture was quenched withice-water and extracted with diethyl ether. The combined extracts werewashed with water, brine and dried over Na₂SO₄ to give ethyl(1-benzyl-piperidin-4-ylidene)cyanoacetate in quantitative yield.R_(f)=0.53 (hexane:AcOEt=1:1).

[B] Synthesis of 1-benzyl-4-cyanomethylpiperidine-4-carbonitrile (3;FIG. 4). A solution of (1-benzyl-piperidin-4-ylidene)cyanoacetate (112.9g, 0.40 mol) in ethanol (500 mL) and water (100 mL), potassium cyanide(64.6 g, 0.99 mol) heated at 65° C. for 24 h. After removal of ethanol,water was added to the residue. The water phase was extracted with etherand the combined ether extract was washed with water, brine and driedover Na2SO4 to give 77.7 g of1-benzyl-4-cyanomethylpiperidine-4-carbonitrile. Rf=0.38(hexane:AcOEt=1:1).

[C] Synthesis of 8-benzyl-2,8-diazaspiro[4,5]decane-1,3-dione (4; FIG.4). 1-Benzyl-4-cyanomethylpiperidine-4-carbonitrile (27.2 g, 0.114 mol),acetic acid (56.8 mL) and sulfuric acid (11.8 mL) were heated at 125° C.for 1 hr. The mixture was cooled to 25° C. and sat. NaOH was used toadjust the pH to 6. The mixture was extracted with dichloromethane. Thecombined extracts were washed with water, brine, dried over Na₂SO₄ andevaporated to give 8-benzyl-2,8-diazaspiro[4,5]decane-1,3-dione.R_(f)=0.40 (CH₂Cl₂:MeOH=10:1).

[D] Synthesis of 8-benzyl-N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione(5; FIG. 4). 8-Benzyl-2,8-diazaspiro[4,5]decane-1,3-dione in DMF wastreated with NaH (1.2 eq) and 2-bromeethane was added. The mixture washeated at 100° C. for 2 hr. The mixture was cooled to 25° C. and pouredover ice-water and extracted with dichloromethane. The combineddichloromethane extract was washed with water, brine, dried over Na₂SO₄and evaporated to give8-benzyl-N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione.

[E] Synthesis of N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione (6; FIG.4). 8-Benzyl-N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione (2.8 g) andPd(OH)₂ (0.8 g) in ethanol (50 mL) and acetic acid (0.5 mL) was stirredunder H₂ at 25° C. for 15 hr. The catalyst was removed by filtration andethanol was evaporated to giveN-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione.

Example 3

One approach to the synthesis of6-O—[N—(N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbonyl]-α,β-D-glucopyranose(13; FIG. 6) is depicted in FIG. 6. 1-O-Benzyl-β-D-Glucoside 7 iscommercially available or it can be made in one step from almondβ-D-glucosidase-catalyzed glycosidation of β-D-glucose and benzylalcohol. See Carbohydrate Res. 1995, 279, 315. In addition, benzyl2,3,4-tri-O-benzyl-6-O-imidazolylcarbonyl-β-D-glucopyranoside (11; FIG.6) can be prepared via a known procedure. See Org. Biomol. Chem. 2003,1, 767-771. Synthetic protocols for the synthesis of the compoundsdepicted in FIG. 6 are presented below.

[A] Synthesis of benzyl 6-O-trityl-β-D-glucopyranside (8; FIG. 6). Asolution of 1 (1.71 g, 6.33 mmol) in pyridine (14 mL) is treated withtrityl chloride (2.65 g, 9.5 mmol) and DMAP (155 mg, 1.27 mmol) at 60°C. for 7 hr. The reaction mixture is cooled at room temperature, MeOH (4mL) added, and the solvents were evaporated. The residue is purified byflash chromatography (hexane-AcOEt, 5:1 to give benzyl6-O-trityl-β-D-glucopyranside as a solid. Rf=0.22 (hexane-AcOEt, 1:2);m.p. 72-74° C.; [α]D²⁰−52.0° (c 1, CHCl₃).

[B] Synthesis of benzyl2,3,4-tri-O-benzyl-6-O-trityl-β-D-glucopyranoside (9; FIG. 6). Asolution of benzyl 6-O-trityl-β-D-glucopyranside (3.58 g, 6.99 mmol) indry DMF (23 mL) is treated with 95% NaH (630 mg, 26.2 mmol) at 0° C.After 10 min, benzyl bromide (2.74 mL, 23.1 mmol) is added and thereaction is allowed to proceed at room temperature overnight. Thereaction mixture is quenched with methanol and concentrated. The residueis dissolved in ether (50 mL), washed with water (4×30 mL), dried(Na₂SO₄), and concentrated. The residue is purified by flashchromatography (hexane-AcOEt, 20:1; 10:1) to give benzyl2,3,4-tri-O-benzyl-6-O-trityl-β-D-glucopyranoside as a syrup. Rf=0.29(hexane-AcOEt, 10:1).

[C] Synthesis of benzyl 2,3,4-tri-O-benzyl-β-D-glucopyranoside (10; FIG.6). A solution of benzyl2,3,4-tri-O-benzyl-6-O-trityl-β-D-glucopyranoside (4.41 g, 5.64 mmol) inCH₂Cl₂-MeOH (1:2, 56.4 mL) is treated with pTsOH (0.21 g, 1.13 mmol) atroom temperature for 6 hr. After this time, triethylamine (0.16 mL, 1.13mmol) is added, the mixture concentrated and the residue dissolved inether (60 mL). The organic solution is washed with water (2×30 mL),brine (30 mL), dried (Na₂SO₄), and concentrated. The residue isfractionated by flash chromatography (hexane-AcOEt, 8:1; 5:1; 3:1) togive benzyl 2,3,4-tri-O-benzyl-β-D-glucopyranoside as a solid.R_(f)=0.21 (hexane-AcOEt, 3:1); m.p. 99-101° C.; [α]D²⁰−10.4° (c 1,CHCl₃).

[D] Synthesis of benzyl2,3,4-tri-O-benzyl-6-O-imidazolylcarbonyl-β-D-glucopyranoside (11; FIG.6). A solution of benzyl 2,3,4-tri-O-benzyl-β-D-glucopyranoside (2.53 g,4.69 mmol) in dioxane (5.3 mL) and N,N-carbonyldiimidazol (0.91 g, 5.63mmol) is stirred at room temperature for 1 hr. After this time, themixture is concentrated and the residue purified by flash chromatography(hexane-AcOEt 3:1) to give benzyl2,3,4-tri-O-benzyl-6-O-imidazolylcarbonyl-β-D-glucopyranoside as asolid. R_(f)=0.22 (hexane-AcOEt, 2:1); m.p. 105-108° C.; [α]D²⁰+28.9° (c1, CHCl₃).

[E] Synthesis of benzyl2,3,4-tri-O-benzyl-6-O—[N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbony]-β-D-glucopyranoside(12; FIG. 6). A mixture of benzyl2,3,4-tri-O-benzyl-6-O-imidazolylcarbonyl-β-D-glucopyranoside (2.78 g,4.39 mmol), N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione (1.63 g, 3.65mmol), triethylamine (5 mL), and THF (15 mL) is stirred at 80° C. for 8hr. After this time, the solvents were removed and the residue waspurified by flash chromatography to give benzyl2,3,4-tri-O-benzyl-6-O—[N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbony]-β-D-glucopyranoside.

[F] Synthesis of6-O—[N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbony]-α,β-D-glucopyranose(13; FIG. 6). A solution of the glycoconjugate (0.01M) inAcOEt-MeOH-toluene (4:3:3) was hydrogenolyzed over 10% Pd/C (1.1 g permmol) for 1-5 hr, when TLC (AcOEt-AcOH-MeOH, 4:1:1) shows the completeconversion product. The reaction mixture is filtered through Celite, andconcentrated. The residue is dissolved in methanol/water (95:5) andfreeze-dried, to give6-O—[N-2-ethyl-2,8-diazaspiro[4,5]decane-1,3-dione)aminocarbony]-α,β-D-glucopyranose.

Example 4

One approach to the synthesis of1,2,3,4,5-penta-O-benzyl-scyllo-inositol is shown in FIG. 7. Preparationof 1,2-O-Cyclohexylidene myo-inositol was done using a modified versionof the method described by Angyal. See J. Chem. Soc. 1961, 4116.1,2,3,4,5,-Penta-O-benzylmyo-inositol was synthesized according to aknown procedure of Lowe. See J. Chem. Soc. Perkin Trans 1. 1991, 1249.Synthetic protocols for the synthesis of the compounds depicted in FIG.7 are presented below.

[A] Synthesis of 1,2-O-cyclohexylidene myo-inositol (2; FIG. 7).Myo-inositol (5 g, 28 mmol) cyclohexanone (50 mL), p-toluene sulfonicacid (36 mg), DMF (5 mL), and benzene (25 mL) were refluxed in aDean-Stark apparatus for 16 hr. The clear solution was cooled to 40° C.and benzene (25 mL), petroleum ether (25 mL), and ethanol (12 mL) wereadded. To this solution was added p-toluene sulfonic acid (0.3 g) andthe mixture was stirred at 4° C. for 2 hr. Triethylamine (0.3 mL) wasadded and the mixture was allowed to stand at −20° C. for 16 hr. Thesuspension was filtered and the filtrate was heated in ethanol (100 mL)and triethylamine (0.5 mL) at 80° C. for 1 hr. After cooling,crystalline (±)-cis-1,2-O-cyclohexylidene myo-inositol was collected byfiltration and dried (5.3 g, 74%), m.p. 175-180° C. (Lit 178° C.).

[B] Synthesis of 1,4,5,6-tetra-O-benzyl-2,3-O-cyclohexylidenemyo-inositol. 1,2-O-cyclohexylidene myo-inositol is treated with benzylbromide and sodium hydride in DMF heated for 16 hr. The solution ispoured over ice-water and extracted with ether. The combined extractsare washed with water, brine and dried. The solvent is evaporated invacuo and the residue is chromatographed on silica gel to give1,4,5,6-tetra-O-benzyl-2,3-O-cyclohexylidene myo-inositol; mp 113-115°C.

[C] Synthesis of 1,4,5,6-Tetra-O-benzylmyo-inositol.1,4,5,6-tetra-O-benzyl-2,3-O-cyclohexylidene myo-inositol is heated for4 hr at 100° C. with glacial acetic acid (20 mL) and water (5 mL). Thesolution is evaporated in vacuo and the residue is chromatographed onsilica gel to give 1,4,5,6-tetra-O-benzylmyo-inositol.

[D] Synthesis of 1,3,4,5,6-Penta-O-benzylmyo-inositol. A solution of1,4,5,6-tetra-O-benzyllmyo-inositol in benzene is treated with benzylbromide and sodium hydride. The mixture was stirred at 100° C. for 2 hr.The reaction mixture is poured over ice-water and extracted with ether.The organic layer was separated and washed successively with saturatedaqueous NaHCO₃ and brine. The solution was evaporated in vacuo and theresidue was chromatographed on silica gel to give1,3,4,5,6-penta-O-benzylmyo-inositol.

[E] Synthesis of 1,2,3,4,5-penta-O-benzyl-scyllo-inositol.1,3,4,5,6-penta-O-benzylmyo-inositol in anhydrous and methylene chloride(20 mL) and dry pyridine is treated with trifluoromethylsulfonylanhydride at −60° C. The reaction mixture is allowed to warm to 25° C.and kept for 2 hr. The solution is quenched with water, diluted withmethylene chloride and washed with saturated aqueous NaHCO₃. Solvent isevaporated in vacuo and the residue is chromatographed on silica gel togive 1,2,3,4,5-penta-O-benzyl-scyllo-inositol.

Example 5

One approach to the synthesis of 1-deoxy-1-[18F]fluoro-scyllo-inositolis presented in FIG. 8. Preparation of1,4,5,6-tetra-O-acetyl-myo-inositol (4; FIG. 8) was done using amodified version of the method described by Angyal. See J. Chem. Soc.1961, 4116. 1,3,4,5,6-Penta-O-acetylmyo-inositol (5; FIG. 8) wassynthesized according to a known procedure of Hosoda. See Bioorg. Med.Chem. 2002, 10, 1855. Synthetic protocols for the synthesis of thecompounds depicted in FIG. 8 are presented below.

[A] Synthesis of 1,2-O-cyclohexylidene myo-inositol (2; FIG. 8).Myo-inositol (5 g, 28 mmol) cyclohexanone (50 mL), p-toluene sulfonicacid (36 mg), DMF (5 mL), and benzene (25 mL) were refluxed in aDean-Stark apparatus for 16 hr. The clear solution was cooled to 40° C.and benzene (25 mL), petroleum ether (25 mL), and ethanol (12 mL) wereadded. To this solution was added p-toluene sulfonic acid (0.3 g) andthe mixture was stirred at 4° C. for 2 hr. Triethylamine (0.3 mL) wasadded and the mixture was allowed to stand at −20° C. for 16 hr. Thesuspension was filtered and the filtrate was heated in ethanol (100 mL)and triethylamine (0.5 mL) at 80° C. for 1 hr. After cooling,crystalline (±)-cis-1,2-O-cyclohexylidene myo-inositol was collected byfiltration and dried (5.3 g, 74%), m.p. 175-180° C. (Lit 178° C.).

[B] Synthesis of 1,4,5,6-tetra-O-acetyl-2,3-O-cyclohexylidenemyo-inositol (3; FIG. 8). 1,2-O-cyclohexylidene myo-inositol (4.8 g,17.4 mmol) was heated at 90° C. for 2 hr with anhydrous pyridine (30 mL)and acetic anhydride (32 mL). The solution was evaporated in vacuo andthe residue was chromatographed on silica gel using 96:4 methylenechloride/methanol to give 1,4,5,6-tetra-O-acetyl-2,3-O-cyclohexylidenemyo-inositol (6.8 g, 85%), mp 112-114° C. (lit 118° C.).

[C] Synthesis of 1,4,5,6-tetra-O-acetylmyo-inositol (4; FIG. 8).1,4,5,6-tetra-O-acetyl-2,3-O-cyclohexylidene myo-inositol (5 g, 11.7mmol) was heated for 4 hr at 100° C. with glacial acetic acid (20 mL)and water (5 mL). The solution was evaporated in vacuo and the residuewas chromatographed on silica gel to give1,3,4,5,6-penta-O-acetylmyo-inositol as a monohydrate.

[D] Synthesis of 1,3,4,5,6-penta-O-acetylmyo-inositol (5; FIG. 8). To asolution of 1,3,4,5,6-penta-O-acetylmyo-inositol (2 g, 5.7 mmol) inanhydrous pyridine (5 mL) and methylene chloride (20 mL) was addedsuccessively 4-(dimethylamino)pyridine (2 mg) and acetyl chloride (0.9g, 11.5 mmol). The mixture was stirred at 25° C. for 16 hr. Water wasadded to the reaction mixture and the organic layer was separated andwashed successively with saturated aqueous NaHCO₃ and brine. Thesolution was evaporated in vacuo and the residue was chromatographed onsilica gel to give 1,3,4,5,6-penta-O-acetylmyo-inositol.

[E] Synthesis of 1,3,4,5,6-penta-O-acetyl-2-O-methanesulfonylmyo-inositol (6; FIG. 8). 1,3,4,5,6-penta-O-acetylmyo-inositol (1 g, 2.6mmol) in anhydrous pyridine (5 mL) and methylene chloride (20 mL) wastreated with 4-(dimethylamino)pyridine (2 mg) and methanesulfonylchloride (0.6 g, 5.2 mmol) at 25° C. for 16 hr. The solution wasevaporated in vacuo and the residue was chromatographed on silica gel togive 1,3,4,5,6-penta-O-acetyl-2-O-methanesulfonyl myo-inositol.

[F] Synthesis of 1-deoxy-1-[18F]fluoro-scyllo-inositol (8; FIG. 8). AWheaton 5-mL reaction vial containing fluorine-18 (100 mCi) in 1 mL of¹⁸O-enriched water, Kryptofix 2.2.2. (8 mg), and potassium carbonate (2mg) was heated at 120° C. and water was evaporated with the aid of anitrogen gas flow. The K¹⁸F/Kryptofix complex was dried three successivetimes by the addition of 1 mL acetonitrile followed by evaporation ofthe solvent using a nitrogen flow. A solution of 2 mg of mesylate 6 in0.1 mL acetonitrile was added to the sealed vial and fluorination wasperformed at 140° C. for 10 min. After cooling to room temperature, thereaction mixture was passed through a silica gel Sep-Pak using methylenechloride (3 mL) and the solvent removed using a nitrogen flow. A mixtureof 0.5 mL 1 M lithium hydroxide and 1 mL methanol was added to thereaction vial and the vial heated at 80° C. for 20 min. Solvent wasremoved, and 1-deoxy-1-[¹⁸F]fluoro-scyllo-inositol was purified on a C18Sep-Pak using saline and filtered (MillexGV 0.22 μm).

Example 6

One approach to the synthesis of 6-O-[(β-D-glucopyranose6-yl)-succinyl]-scyllo-inositol is shown in FIG. 9. Benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside 6-(hydrogen succinate) isprepared from a known procedure. See Org. Biomol. Chem. 2003, 1,767-771. Synthetic protocols for the synthesis of the compounds depictedin FIG. 9 are presented below.

[A] Synthesis of benzyl 2,3,4-tri-O-benzyl-β-D-glucopyranoside6-(hydrogen succinate). A solution of benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside (10; FIG. 9) in pyridine istreated with succinic anhydride and DMAP and stirred at room temperaturefor 16 h. The mixture is concentrated and the residue dissolved inCH₂Cl₂ and washed successively with 5% aqueous HCl (3×15 mL), sat.aqueous NaHCO₃ (3×15 mL), and water (3×15 mL). The organic solution isdried (Na₂SO₄), and concentrated to give benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside 6-(hydrogen succinate) as asolid. R_(f)=0.56 (hexane-AcOEt, 2:1); m.p. 75-80° C.; [α]D²⁰−7.7° (c 1,CHCl₃).

[B] Synthesis of benzyl-[(benzyl 2,3,4-tri-O-benzyl-β-D-glucopyranoside6-yl)-succinyl]-scyllo-inositol. A solution of benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside 6-(hydrogen succinate) in CH₂Cl₂(2.5 mL) is added dropwise to a solution of1,2,3,4,5,-penta-O-benzyl-scyllo-inositol andN,N′-diisopropylcarbodiimide in CH₂Cl₂ (10 ml) at 0° C. The mixture isallowed to stir 24 h at room temperature, then, diluted with CH₂Cl₂ (15mL), washed with 1N KHSO₄ (2×10 mL), sat. aqueous NaHCO₃ (2×10 mL),water (2×10 mL), and sat. NaCl (10 mL). The organic solution is dried(Na₂SO₄), the solvent removed under reduced pressure, and the residuepurified by flash chromatography to give benzyl-[(benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside 6-yl)-succinyl]-scyllo-inositol.

[C] Synthesis of 6-O-[(β-D-glucopyranose6-yl)-succinyl]-scyllo-inositol. Benzyl-[(benzyl2,3,4-tri-O-benzyl-β-D-glucopyranoside 6-yl)-succinyl]-scyllo-inositolin 1:1 AcOEt-MeOH is treated with TFA and Pd/C and stirring underhydrogen for 12 h. The reaction mixture is filtered and concentrated,and the residue is dissolved in CH₃NO₂-water (95:5) and freeze-dried, togive 6-O—[(β-D-glucopyranose 6-yl)-succinyl]-scyllo-inositol.

Example 7

RS-86 (2-ethyl-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dione), a M1muscarinic receptor agonist, was radiofluorinated and brain accumulationevaluated and compared to that of its C-11 labeled analog. Alzheimer'sdisease (AD) is associated with reductions in the presynaptic markercholine acetyltransferase activity and M2 muscarine receptors whichprecedes other pathologic changes. A number of M1 agonists have beendeveloped to treat AD that increase acetylcholine transmission in thebrain alleviating memory loss. Ml postsynaptic receptors are thought tobe upregulated. Therefore, a M1 specific radioligand may show higherthan normal accumulation in patients, and hence, serve as a potentialearly diagnostic method for AD.

As described above,2-(2-methanesulfonylethyl)-8-methyl-2,8-diazaspiro[4,5]decane-1,3-dionewas prepared in five steps. The RS-86 mesylate (2 mg) in acetonitrile(0.1 mL) was added to a sealed vial containing dried K¹⁸F/kryptofix andheated at 120° C. for 10 min. The mixture was diluted with water (0.2mL) and purified by HPLC (C-18 column 250×10 mm, flow 6 mL/min,PBS/acetonitrile (95:5). The fraction containing [¹⁸F]fluoro-RS 86 wasevaporated to dryness and the activity dissolved in PBS beforefiltering. Biodistribution was performed in rats at 5, 30 and 60 min.

[¹⁸F]fluoro-RS 86 was prepared in 20% yield (EOS) with 95% chemical and98% radio purity. Brain uptake at 5, 30 and 60 min was 0.27, 0.54, 0.48,respectively. Blood, liver and stomach were 0.3%, 0.64% and 0.90% withthe greatest uptake in the kidneys (2.5%) at 30 min. Defluorination wasminimal (0.42% in bone at 30 min).

The lower brain uptake of [¹⁸F]fluoro-RS 86 compared to [¹¹C]RS-86(0.54% vs. 1%) may be due to the location of the fluoride label on theligand. These preliminary results suggest that [¹⁸F]fluoro-RS 86accumulates in rat brain, however further studies are needed todetermine whether the distribution in the brain reflects M1 mAChRconcentration.

INCORPORATION BY REFERENCE

All of the U.S. patents and U.S. patent application publications citedherein are hereby incorporated by reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A compound, or a pharmaceutically acceptable salt thereof,represented by formula II:

wherein, independently for each occurrence, L is —H, —C(═O)R³,—C(═O)[C(R¹)₂]_(p)R³, —C(═O)[C(R¹)₂]_(p)C(═O)R³, —[C(R¹)₂]_(p)C(═O)R³,—[C(R¹)₂]_(p)R³, —[C₁₋₁₀alkylene]R³, —C(═O)[C₁₋₁₀alkylene]R³,—[C₁₋₁₀alkylene]C(═O)R³ or —C(═O)[C₁₋₁₀alkylene]C(═O)R³; R¹ is hydrogen,halo, azido, alkyl, haloalkyl, perhaloalkyl, fluoroalkyl,perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy, aryloxy,heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano, orisocyano;

R⁴ is hydrogen, halo, azido, alkyl, haloalkyl, perhaloalkyl,fluoroalkyl, perfluoroalkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy,aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino,arylamino, acylamino, heteroarylamino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl,thioether, sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano,isocyano, —OR⁵, —SR⁵, —N(R⁵)₂, —(C(R¹)₂)_(q)C(R¹)₃ or a chelatingstructure; R⁵ is hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, acyl,—(C(R¹)₂)_(q)C(R¹)₃ or a chelating structure; R⁷ is hydrogen, halo,azido, alkyl, haloalkyl, perhaloalkyl, fluoroalkyl, perfluoroalkyl,aralkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,aralkyl, heteroaralkyl, hydroxy, alkoxy, aryloxy, heteroaryloxy,aralkyloxy, heteroaralkyloxy, amino, alkylamino, arylamino, acylamino,heteroarylamino, nitro, sulfhydryl, imino, amido, phosphonate,phosphinate, acyl, carboxyl, oxycarbonyl, acyloxy, silyl, thioether,sulfo, sulfonate, sulfonyl, sulfonamido, formyl, cyano, isocyano, —OR⁵,—SR⁵, —N(R⁵)₂, —(C(R¹)₂)_(q)C(R¹)₃ or a chelating structure; p is 1-10inclusive; and q is 0-10 inclusive; provided that when L is —H, one R⁷is fluoro; and the other R⁷ are hydroxy, alkoxy, aryloxy, heteroaryloxy,aralkyloxy, or heteroaralkyloxy.
 2. The compound of claim 1, whereinsaid compound comprises at least one ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I or¹²⁵I.
 3. The compound of claim 1, wherein said compound comprises atleast one ¹⁸F.
 4. The compound of claim 1, wherein L is —C(═O)R³ or—C(═O)[C(R¹)₂]_(p)C(═O)R³.
 5. The compound of claim 1, wherein L is—C(═O)R³.
 6. The compound of claim 1, wherein L is—C(═O)[CH₂]_(p)C(═O)R³.
 7. The compound of claim 1, wherein L is—C(═O)CH₂CH₂C(═O)R³.
 8. The compound of claim 1, wherein R⁷ is —H, —F,—Cl, —Br, —I, or —OH.
 9. The compound of claim 1, wherein R⁷ is —F or—OH.
 10. The compound of claim 1, wherein R⁷ is —¹⁸F or —OH; providedthat only one R⁷ is —¹⁸F.
 11. The compound of claim 1, wherein R³ is


12. The compound of claim 1, R³ is

and R⁴ is —H, —F, —Cl, —Br, —I, or —OR⁵.
 13. The compound of claim 1, R³is

and R⁴ is —OH.
 14. The compound of claim 1, wherein the compoundcomprises a radioimaging agent.
 15. The compound of claim 1, wherein Lis —H, one R⁷ is fluoro; and the other R⁷ are hydroxy, alkoxy, aryloxy,heteroaryloxy, aralkyloxy, or heteroaralkyloxy.
 16. The compound ofclaim 1, wherein the compound is 1-deoxy-1-fluoro-scyllo-inostiol or1-deoxy-1-fluoro-myo-inositol.
 17. The compound of claim 14, wherein R¹,R⁴, or R⁷ comprises the radioimaging agent; and the radioimaging agentis ¹⁸F.
 18. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a compound of claim
 1. 19. A method for amyloidimaging a subject suffering from an amyloidosis-associated pathologicalcondition comprising the step of: administering a diagnosticallyeffective amount of a compound of claim
 1. 20. The method of claim 19,wherein said amyloidosis-associated pathological condition isAlzheimer's disease.
 21. The method of claim 19, wherein said compoundcomprises at least one ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I or ¹²⁵I.
 22. Themethod of claim 19, wherein the compound is


23. A method for treating a subject suffering from anamyloidosis-associated pathological condition, comprising the step of:administering a therapeutically effective amount of a compound ofclaim
 1. 24. The method of claim 23, wherein said amyloidosis-associatedpathological condition is Alzheimer's disease.
 25. The method of claim23, wherein said compound is a combination of inositol and glucose, or adisaccharide comprising inositol and glucose.
 26. The method of claim23, wherein said compound is fluorinated.